Transport of Milk Constituents by The Mammary Gland

PHYSIOLOGICAL REVIEWS
Vol. 80, No. 3, July 2000

Printed in U.S.A.
Transport of Milk Constituents by the Mammary Gland

D. B. SHENNAN AND M. PEAKER
Hannah Research Institute, Ayr, Scotland
I. Introduction 926
II. Experimental Methods Used to Study Mammary Gland Solute Transport 926
III. Routes of Secretion 927
A. Description of routes 927
B. Solute transport via the paracellular pathway 928
IV. Sodium, Potassium, and Chloride Transport 930
A. Carrier mechanisms for Na⫹, K⫹, and Cl⫺ 930
B. Na⫹, K⫹, and Cl⫺ channels in the apical membrane 931
C. Effects of prolactin on Na⫹, K⫹, and Cl⫺ transport 931
V. Relations Between Sodium, Potassium, and Chloride Transport and Water Movements 932
VI. Phosphate Transport 932
VII. Calcium Transport 933
VIII. Citrate Transport 934
IX. Iodide Transport 934
X. Stretch-Sensitive Ion Transport Mechanisms 934
XI. Choline and Carnitine Transport 935
XII. Glucose Transport 935
A. Glucose transporters 935
B. D-Glucose transport across the apical membrane 936
C. Transport of D-glucose across the Golgi membrane 937
D. Control of D-glucose transport 937
XIII. Amino Acid Transport 938
A. Amino acid transporters 938
B. Amino acid transport across the apical membrane of mammary secretory cells 941
C. Control of amino acid transport 941
D. Volume-sensitive amino acid transport 942
E. Peptide transport 943
F. Does ␥-glutamyl transpeptidase play a role in mammary amino acid transport? 944
XIV. Fatty Acid Uptake 945
XV. Coordination and Control 945
Shennan, D. B., and M. Peaker. Transport of Milk Constituents by the Mammary Gland. Physiol Rev 80: 925–951,
2000.—This review deals with the cellular mechanisms that transport milk constituents or the precursors of milk
constituents into, out of, and across the mammary secretory cell. The various milk constituents are secreted by
different intracellular routes, and these are outlined, including the paracellular pathway between interstitial fluid and
milk that is present in some physiological states and in some species throughout lactation. Also considered are the
in vivo and in vitro methods used to study mammary transport and secretory mechanisms. The main part of the
review addresses the mechanisms responsible for uptake across the basolateral cell membrane and, in some cases,
for transport into the Golgi apparatus and for movement across the apical membrane of sodium, potassium, chloride,
water, phosphate, calcium, citrate, iodide, choline, carnitine, glucose, amino acids and peptides, and fatty acids.
Recent work on the control of these processes, by volume-sensitive mechanisms for example, is emphasized. The
review points out where future work is needed to gain an overall view of milk secretion, for example, in marsupials
where milk composition changes markedly during development of the young, and particularly on the intracellular
coordination of the transport processes that result in the production of milk of relatively constant composition at
a particular stage of lactation in both placental and marsupial mammals.
www.physrev.physiology.org 0031-9333/00 $15.00 Copyright © 2000 the American Physiological Society 925
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926 D. B. SHENNAN AND M. PEAKER Volume 80
I. INTRODUCTION various mammary gland preparations, effects on trans-

port may be observed. However, it is difficult to determine
The last Physiological Reviews article on the mech- whether a hormone is having a primary, direct effect on a
anism of milk secretion was written in 1971 (118) at a time particular transport system or whether the effect is due to
when morphological information from electron micros- the overall response of the cell, to increase the degree of
copy was being combined with physiological and bio- differentiation, for example, or simply to prolong its sur-
chemical knowledge to provide the basis of our current vival. Therefore, caution must be exercised in interpreting
understanding of milk secretion at the cellular level. Many the individual effects of hormones on transport systems in
of the hypotheses on the secretory mechanisms for the such highly integrated differentiation, secretion, and
various milk constituents have survived more extensive transport systems.
work and amplification at the cellular and molecular lev-
els, and it is not our intention to cover this older ground
other than in outline. Rather, our intention is to extend II. EXPERIMENTAL METHODS USED TO STUDY
the explanatory reductionist approach to examine the MAMMARY GLAND SOLUTE TRANSPORT
transport and secretory mechanisms for the major milk
components or their precursors and their integration The first physiological studies on the mechanism of
within the economy of the secretory cell. milk secretion were done using conscious undisturbed
The mammary gland is, in a number of respects, an animals. Although the range of measurements that can be
unusual exocrine gland. Secretion, once initiated at about made is limited, the major advantage is that artifacts are
the time of parturition, is continuous but relatively slow in not introduced as part of the experimental procedure of
terms of fluid volume output per unit time. The secretion, removing tissue and trying to maintain it alive and undam-
milk in full lactation, is a complex mixture arising from aged. It is relatively easy to measure transepithelial solute
different secretory pathways across, within, and some- gradients across the lactating mammary gland, and more-
times between the mammary epithelial cells, comprising over, because of the slow rates of milk secretion, it is
membrane-bound fat droplets, casein micelles, and an possible to make some meaningful electrophysiological
aqueous phase that usually contains lactose and some- measurements; the effect of changing the luminal con-
times complex carbohydrates, minerals in various ionic or tents on the transepithelial potential difference can be
bound forms, other proteins, and a bewildering array of readily tested, for example (27).
soluble components, some of which may be biologically Many studies on the mechanism of solute transport
active in the young consuming the milk (78, 165). Another have been done using mammary tissue slices, otherwise
but related unusual feature is that secretion is stored termed fragments and explants. Explants are still a pop-
within the lumen of the gland, either in the secretory ular choice because they are easily and rapidly prepared;
alveoli or duct system, until it is removed at intervals they do not require biochemical treatment, such as colla-
(varying from several hours in many mammals, to once genase digestion, which could possibly alter the proper-
daily in rabbits, to once every 2 days in tree shrews, and ties of the alveolar cells. Although explants are a mixed
to once a week in some pinnipeds) by the sucking young cell population, the vast majority, at least in those pre-
(46). pared from lactating tissue, are secretory cells. The alve-
Studies in a number of species have shown that while olar nature of mammary tissue means that explants will
there may be species-specific changes in milk composi- give a good measure of transport across the basolateral
tion with stage of lactation corresponding to growth membranes of the secretory cells particularly in experi-
phases of the young (a phenomenon particularly marked ments employing short time courses. However, the rela-
in marsupials), milk composition is markedly resistant to tively large extracellular space of mammary tissue ex-
environmental, including nutritional, changes, whereas plants together with the presence of large unstirred fluid
the quantity of milk secreted shows considerable plastic- layers places limitations on the design of experiments.
ity (162). Isolated mammary epithelial cells and acini are an alter-
The control of the rate of milk secretion will not be native to explants to study membrane transport. Like
covered in detail, except for the effects of prolactin and explants, they are relatively easy to prepare; however, the
cell stretch, since it has been reviewed extensively in isolation process requires that the cells be exposed to
recent years, particularly the autocrine control of milk digestive agents. The extracellular space is minimal com-
secretion that acts to match supply by the mother to pared with that associated with explants, but it is difficult
demand by the young within each individual mammary to distinguish between transport occurring at the apical
gland within the hormonal milieu of lactation that main- and basolateral membranes in both these preparations.
tains the cells in a secretory state (163, 168, 169, 247–249). There is also the possibility that dissociation of mammary
The mammary gland is controlled by a number of tissue leads not only to a loss of polarity but also de-
hormones and local factors. When these are added to creases cell viability.

July 2000 MAMMARY TRANSPORT MECHANISMS 927
The perfused lactating mammary gland has been suc-

cessfully employed to characterize solute transport sys-
tems in both guinea pig and rat. This technique allows the
transport properties of the blood-facing aspect of the
mammary epithelium to be characterized under near-
physiological conditions. One major assumption is that
the endothelium does not constitute a major barrier to
solute movement between blood and the interstitium.
Recent work suggests that this is a reasonable assump-
tion, since sucrose (an extracellular marker) has a larger
dilution space than albumin (an intravascular marker) in
the lactating bovine mammary gland (176). The major
drawback is that the skills required to achieve a success-
ful preparation take time to acquire. The perfused mam-
mary gland, used in conjunction with the rapid, paired-
tracer dilution technique (255), has been successfully
employed to study amino acid transport.
Cultured mammary cells have also been used to
⫹
study mammary transport processes. For example, cells FIG. 1. K (Rb⫹) uptake by milk-fat-globule membrane vesicles
cultured on collagen gels to form monolayers and subse- (squares) and skim milk membrane vesicles (circles) prepared from
bovine milk in the absence (open symbols) and presence (solid symbols)
quently mounted in Ussing chambers have allowed the of valinomycin. Previously, it has been argued that both of these mem-
polarity of the mammary epithelium to be examined. brane preparations originate from the apical aspect of mammary secre-
However, there is justifiable concern that cultured mam- tory cells. It is apparent that skim milk membrane vesicles are perme-
able to K⫹ in the absence and presence of valinomycin. In contrast,
mary epithelial cells are not fully active in terms of secre- milk-fat-globule membranes only display a significant permeability to K⫹
tion, although an exception to this may be mammary in the presence of valinomycin. Data are means ⫾ SE (n ⫽ 3). The
epithelial cells cultured on a reconstituted basement results suggest that skim milk membranes may be a better marker for
the apical membrane of mammary secretory cells than milk-fat-globule
membrane. Cells grown in this manner form polarized membranes. [Adapted from Shennan (195).]
acinar structures that enclose a hollow, sealed lumen and
secrete in a vectorial fashion (13, 42, 89). These acini,
termed mammospheres, may prove particularly useful for these membranes (195, 196) (Fig. 1). There is the possi-
characterizing transport across the basolateral mem- bility that the milk-fat-globule membrane is derived from
branes of secretory cells. Unfortunately, a technique that intracellular membranes rather than the apical membrane
allows purified basolateral membrane vesicles to be pre- (228, 253). On the other hand, membrane vesicles pre-
pared from mammary secretory cells has not yet been pared from bovine skim milk, which are distinct from
developed. milk-fat-globule membranes, are able to transport solutes
Membrane fractions isolated from milk have been into an intravesicular compartment (196). Skim milk
used in attempts to study the transport properties of the membrane vesicles do appear to originate from the apical
apical (luminal) membrane. Wooding et al. (254) identi- membrane (172).
fied membrane-bound cytoplasmic droplets in goat’s milk In ending this section, it should be noted that the
containing fat and protein particles, mitochondria, and relatively complex morphology of the mammary gland
Golgi apparatus (but no nuclei); further studies showed precludes the use of several techniques to study solute
that the droplets were capable of triacylglycerol biosyn- transport. For example, the mammary epithelium is not
thesis (44). Because the cytoplasmic fragments originate flat and therefore, unlike frog skin and toad bladder,
from the apical aspect of mammary secretory cells, it is cannot be mounted in Ussing chambers.
reasonable to assume that the surrounding membrane is
indeed representative of the apical membrane. It has been
III. ROUTES OF SECRETION
shown that cytoplasmic droplets transport ions into an
osmotically active space (213).
Because the milk fat globule becomes enveloped by A. Description of Routes
membrane during its extrusion from the cell, the milk-fat-
globule membrane has long been assumed to be repre- There are five major, known routes of secretion
sentative of the apical membrane of the secretory cell. across the mammary secretory epithelium from the blood
However, the finding that bovine milk-fat-globule mem- side to milk, four transcellular and one paracellular (116,
brane vesicles are impermeable to K⫹ casts doubt on the 118) (Fig. 2): 1) membrane route, 2) Golgi route, 3) milk
accepted origin or maintenance of original properties of fat route, 4) transcytosis, and 5) paracellular route.

FIG. 2. Diagram showing the known major

routes of secretion across the mammary epithe-
lium. Routes 1, 2, 3, and 5 correspond to the de-
scriptions given in text. For the sake of clarity,
route 4 (transcytosis) is not shown. [Adapted from
Linzell and Peaker (118).]
1) In the membrane route, substances may traverse 4) In transcytosis, vesicular transport involves vari-
the apical cell membrane (and for those directly derived ous organelles, possibly, in some cases also involving
from blood, the basolateral membrane). Examples are extrusion by route 2 (88, 154). Examples are immuno-
water, urea, glucose, Na⫹, K⫹, and Cl⫺ (116). globulins during colostrum formation (88), transferrin
2) In the Golgi route, secretory products are trans- (154), and prolactin (154).
ported to or sequestered by the Golgi apparatus and se- 5) In the paracellular route, there is direct passage
creted into the milk space by exocytosis. Examples are from interstitial fluid to milk.
casein, whey proteins, lactose, citrate, and calcium (116, In full lactation, routes 1, 2, 3, and, possibly, 4 pre-
118, 267). dominate.
3) In the milk fat route, milk fat globules are ex-
truded from the apex of the secretory cell surrounded by
membrane (milk-fat-globule membrane); some cytoplasm B. Solute Transport Via the Paracellular Pathway
is sometimes included (126). Examples are milk fat, lipid-
soluble hormones and drugs, some unknown growth fac- Transcellular transport requires solutes to cross the
tors (in milk-fat-globule membrane) (244), and the prod- constituent plasma membranes of the epithelium,
uct of the ob gene, leptin (214). whereas paracellular transfer involves movement be-

tween the cells via leaky tight junctions. In most species, transepithelial electrical resistance and decreased the
solute transport across the lactating mammary epithelium transepithelial movements of labeled mannitol and inulin
appears to be mainly transcellular, i.e., the zonulae occlu- across monolayers of a mouse mammary cell line (261).
dentes are tight junctions. However, there are situations Glucocorticoid-induced formation of mouse mammary
where a significant flux occurs via a paracellular pathway. tight junctions involves an increase in the amount of the
Movement of solutes across the paracellular shunt path- tight junction-associated protein ZO-1 and phosphoryla-
way occurs during pregnancy and, in some species, dur- tion/dephosphorylation events since okadaic acid, a pro-
ing late lactation. The early (160) and later work (150) on tein serine/threonine phosphatase inhibitor, attenuates
this aspect of mammary transport including physiological the dexamethasone-induced increase in transepithelial re-
and morphological findings have been reviewed, and it is sistance (212). However, mammary development is a
not our intention to provide a further detailed review complex series of events, and transforming growth factor
here. (TGF)-␤ was found to antagonize the effect of dexameth-
The transepithelial electrical resistance is a good asone (252). There is evidence from in vivo studies that
measure of the tightness (or leakiness) of an epithelium. glucocorticoids may control the permeability of tight
However, although it is possible to measure the transep- junctions. In late-pregnant goats, an intraluminal injection
ithelial resistance of cultured mammary cell monolayers, of cortisol had marked effects on the composition of fluid
it is more difficult to obtain reliable values of the electri- within the lumen: K⫹ concentration increased and Na⫹
cal resistance across the lactating mammary gland in vivo. decreased, which is consistent with the closing of tight
A common way of assessing paracellular permeability in junctions (224). However, an effect of cortisol on trans-
the lactating mammary gland is to measure the solute cellular ion transport cannot be ruled out at this stage.
content of milk; the concentrations of lactose and K⫹ in Hormonal events, other than or as well as a rise in circu-
milk decrease (moving down their concentration gradi- lating glucocorticoid concentration, appear to be involved
ents), whereas those of Na⫹ and Cl⫺ increase when the in the final closure near parturition, with the hormonal
junctions become leaky. However, care must be exercised events leading to parturition and the onset of copious
when relying solely on Na⫹ and K⫹ gradients because a milk secretion (lactogenesis stage II, Ref. 68) being
change may simply reflect an alteration in the transport of closely linked. For example, in late pregnancy, the goat
these ions across transcellular pathways. A more prefer- mammary gland synthesizes PGF2␣; near term, this pro-
able way of assessing tight junction permeability in vivo is duction stops. Unilateral treatment with a PGF2␣ analog
to measure the rate of appearance of lactose in plasma or prevented the normal changes in milk composition and
of an inert disaccharide introduced into plasma in milk increase in the rate of secretion (127). The current view
(119, 216). on the control of tight junctional permeability during
In general, the paracellular pathway is absent during lactogenesis has been well summarized by Nguyen and
lactation and present during pregnancy. However, in the Neville (150)
rabbit during late lactation, when the rate of secretion is
still relatively high, milk lactose and K⫹ concentrations We propose that progesterone inhibits tight junc-
decrease while those of Na⫹ and Cl⫺ increase. At this tion closure during pregnancy, possibly through or
stage, the epithelium is permeable to lactose and sucrose along with local factors such as TGF-␤ and PGF2␣. As
the level of progesterone drops during parturition or
and more permeable to Na⫹ and Cl⫺ (167). These changes with ovariectomy, its inhibitory effect is removed,
were reversed by exogenous prolactin (121). Recent evi- allowing the mammary epithelial tight junctions to
dence suggests that prolactin may also be involved in close. However, glucocorticoid may be required for
some way in maintaining the integrity of the tight junc- final differentiation of the mammary epithelium from
tions (judged by the ability of the gland to maintain solute the pregnant animal into the epithelium of the lactat-
ing animal with its highly impermeable tight junctions.
concentration gradients) in the rat mammary gland (71). Prolactin appears to play a role in the maintenance of
There is also evidence that a paracellular pathway ap- the mammary epithelium in the lactating state and
pears during other physiological states. Characteristic inhibition of programmed cell death.
changes in milk composition were observed in many
goats for 1–2 days up to 4 days before estrus (164), but the The paracellular pathway reappears at the cessation
controlling mechanism remains unknown. of lactation. When milk removal was stopped in goats
There is a major change in the permeability of the during late lactation, the key process was determined to
paracellular pathway at the onset of copious milk secre- be the local event of stretching the mammary epithelium
tion around the time of parturition (119). There is also by milk accumulating within the lumen of the gland,
good evidence which suggests that glucocorticoids are leading to a decrease in secretory and metabolic activity
involved in controlling the formation of tight junctions and an increase in the permeability of the tight junctions.
during mammary development and differentiation in preg- Systemic control by hormones was excluded in this spe-
nancy. The glucocorticoid dexamethasone increased the cies as was pressure of the accumulated milk simply

bursting the taut epithelium (69, 161). Similarly, in the

cow, milk stasis leads to an increase in the paracellular
pathway that can be reversed by milking (217). These as
well as other studies indicate an inverse link between
secretory activity and permeability of the tight junctions.
Whether maintenance of impermeable junctions is part of
the secretory activity of the cell, like the synthesis of milk
components, or whether disruption of the junctions is
followed by partial depolarization of the cell which affects
secretory activity, or whether there is signaling from the
junctions to the synthetic and secretory pathways is not
known (150).
The presence of a paracellular pathway may con-
found studies on milk composition and mammary epithe-
lial permeability. Disruption of the epithelium is evident
in mastitis but also when supraphysiological doses of
oxytocin are given to contract the myoepithelium and
obtain a milk sample. Care must be exercised when inter-
preting data on milk composition, particularly of the aque-
ous phase (160).
IV. SODIUM, POTASSIUM, AND CHLORIDE

TRANSPORT
The mammary gland is able to generate and maintain

large Na⫹, K⫹, and Cl⫺ gradients between milk and
plasma. These ions make a substantial contribution to the
osmolality of milk, especially in those species where the
lactose content is relatively low. On the basis of measure-
ments of the concentration gradients between interstitial
or extracellular fluid, intracellular fluid, and milk, and the
FIG. 3. Profile of ion distribution and membrane potentials (both
electrical potential gradients between these three com- measured and calculated) in guinea pig mammary tissue. [Modified from
partments, Linzell and Peaker in 1971 (117, 118) proposed Linzell and Peaker (118).]
a scheme to account for the movement of Na⫹, K⫹, and
Cl⫺ across the lactating mammary epithelium and the
maintenance of milk composition (Fig. 3). In their hypoth- concentration, apical membrane potential difference, and
esis, the main features are that the Na⫹ concentration ionic concentrations have also been considered (160,
inside the cell is kept low, and the K⫹ concentration high, 159).
by the action of a Na⫹-K⫹ pump on the basolateral mem-
brane, whereas these ions are freely distributed between
the interior of the cell and milk according to the electrical A. Carrier Mechanisms for Naⴙ, Kⴙ, and Clⴚ
potential difference across the apical membrane. Milk is
electrically positive with respect to the cell, so the con- Three carrier mechanism were postulated in the
centrations of Na⫹ and K⫹ are lower in milk, but the ratio scheme of Linzell and Peaker: 1) a Na⫹-K⫹ pump on the
of these ions is similar in both compartments, i.e., ⬃3 basolateral membrane, 2) an inwardly directed Cl⫺ pump
K⫹:1 Na⫹. The situation for Cl⫺ is, however, different on the apical membrane, and 3) a similar Cl⫺ transporter
from that of Na⫹ and K⫹. The concentration of Cl⫺ is in the basolateral membrane of the secretory cell (117,
higher inside mammary cells than that expected for an 118). Evidence for the presence of a Na⫹-K⫹ pump in the
equilibrium distribution, suggesting that a mechanism ex- blood-facing aspect of the mammary epithelium is strong,
ists in both the basolateral and apical membranes which and several candidates for Cl⫺ pumps have been identi-
“actively” pumps Cl⫺ into the cells. Many experimental fied.
observations, at the level of individual transport systems, Many studies have shown that lactating mammary
supporting the original hypothesis of Linzell and Peaker tissue from a variety of species displays ouabain-sensitive
have since been made. The relations between lactose Na⫹-K⫹-ATPase activity (242) and ouabain-sensitive Na⫹

and K⫹ transport (3, 61, 62, 117, 194). In addition, there is ple, a Cl⫺/HCO⫺3 antiport drives the uphill accumulation
good histochemical evidence that Na⫹-K⫹-ATPase is lo- of Cl⫺ into smooth muscle cells (2, 50). A DIDS-sensitive
cated on the basal and lateral membranes, but not the anion exchanger that accepts Cl⫺, SO2⫺ ⫺ ⫺
4 , I , and NO3 as
apical membranes, of mammary secretory cells (96, 105). substrates has been reported in lactating rat mammary
These findings taken together support the scheme of Lin- tissue explants (194, 203). However, it remains to be
zell and Peaker that lactating mammary tissue expresses shown whether the rat mammary tissue anion-exchanger
a functional Na⫹-K⫹ pump on the blood-facing pole of the transports HCO⫺3 and, if so, whether it is able to act as a
epithelium. Experiments using cultured mammary epithe- Cl⫺ pump.
lial cells have provided evidence that the Na⫹-K⫹ pump is
located on the basolateral, but not the apical, membranes
B. Naⴙ, Kⴙ, and Clⴚ Channels in the
(24 –26). However, there is evidence for Na⫹-K⫹-ATPase
Apical Membrane
activity on the milk-fat-globule membrane, albeit at a
relatively low specific activity (55).
The 1971 model of Linzell and Peaker predicts that
Na⫹-K⫹-Cl⫺ cotransport could be a candidate for the
the apical membrane of mammary secretory cells pos-
Cl pump in the model of Linzell and Peaker. Na⫹-K⫹-Cl⫺
⫺
sesses Na⫹, K⫹, and Cl⫺ channels. Several lines of exper-
cotransport has been identified in lactating rat mammary
imental evidence support this prediction. First, Blatch-
tissue (194, 203). Thus K⫹ (Rb⫹) transport (both influx
ford and Peaker (27) have shown that the transepithelial
and efflux) by mammary tissue explants is Na⫹ depen-
potential difference across the lactating goat mammary
dent, Cl⫺ dependent, and inhibited by the loop diuretic
gland can be altered by varying the Na⫹, K⫹, and/or Cl⫺
furosemide. In addition, Cl⫺ efflux from rat mammary
concentration in the lumen of the gland in a fashion that
tissue explants is sensitive to furosemide (203). On the
is consistent with the presence of channels for each of
basis that cotransport activity was detected in explants, it
these ions in the apical aspect. However, it was found that
is predicted that the Na⫹-K⫹-Cl⫺ cotransport system is
the apical membrane is unable to discriminate between
situated in the blood-facing aspect of the epithelium. Sup-
Na⫹ and K⫹, suggesting that the apical membrane may
porting evidence of Na⫹-K⫹-Cl⫺ expression in mammary
express a nonspecific cation channel rather than separate
tissue is the finding that bovine milk-fat-globule mem-
channels for Na⫹ and K⫹. In this connection, nonspecific
brane binds bumetanide (an analog of furosemide) in a
cation channels have been identified in cultured mouse
fashion that is dependent on Na⫹, K⫹, and Cl⫺ (166).
mammary epithelial cells (72). Second, cytoplasmic frag-
The triple cotransporter is able to drive the uphill
ments isolated from goat’s milk that are surrounded by
transport of Cl⫺ (2). The Na⫹-K⫹-Cl⫺ cotransport system
apical membrane, transport K⫹(Rb⫹) via barium- and qui-
is electroneutral and is generally assumed to operate with
nine-sensitive channels (213). Third, skim milk mem-
a stoichiometry of 1 Na⫹:1 K⫹:2 Cl⫺ (43, 79) The free-
branes, isolated from bovine milk, which appear to be
energy (⌬G) available to a cotransporter operating with
derived from the apical aspect of secretory cells, express
such a stoichiometry is given by
K⫹ and Cl⫺ channels (196). The channels identified in the
membrane preparations need to be characterized using
⌬G ⫽ RT ln 冉关Na ⫹ 兴 i 关K ⫹ 兴 i 关Cl ⫺ 兴 2i
关Na ⫹ 兴 o 关K ⫹ 兴 o 关Cl ⫺ 兴 o2 冊 (1)
electrophysiological techniques.
C. Effects of Prolactin on Naⴙ, Kⴙ, and

where subscripts i and o refer to intracellular and extra- Clⴚ Transport
cellular, respectively. Applying the Na⫹, K⫹, and Cl⫺ con-
centrations of guinea pig mammary tissue and plasma Prolactin was found to decrease the Na⫹ and in-
(117) to Equation 1 gives a value ⫹0.79RT for G, suggest- crease the K⫹ content of mammary tissue taken from
ing that the triple cotransport system operating with a rabbits treated with bromocriptine, a drug that prevents
coupling ratio of 1 Na⫹:1 K⫹:2 Cl⫺ would require an input prolactin release; ouabain inhibited the action (62, 63). In
of free energy to drive inward transport. Alternatively, a addition, the unidirectional uptake of K⫹(Rb⫹) by rabbit
Na⫹-K⫹-Cl⫺ cotransporter with a stoichiometry of 2 mammary explants was stimulated by prolactin via a path-
Na⫹:1 K⫹:3 Cl⫺, as found in the squid axon (190), would way sensitive to ouabain (61). These observations are
have sufficient freeenergy (G ⫽ ⫺1.03RT) associated with consistent with an effect of prolactin-induced differentia-
the ion gradients to accomplish inward transport. How- tion of the cells on the number of Na⫹-K⫹ pumps present
ever, it is clear that further experiments are required to either induced directly or through an increase in Na⫹
determine both the stoichiometry and locus of the mam- permeability. In this connection it is interesting to note
mary Na⫹-K⫹-Cl⫺ cotransport mechanism. that prolactin upregulates Na⫹-K⫹-Cl⫺ cotransport in rat
Cl⫺/HCO⫺ 3 exchange could also be a candidate for a mammary tissue (202). Work using cultured cells sup-
⫺
Cl pump in lactating mammary tissue (120). For exam- ports the claim that prolactin-induced differentiation in-

volves changes in mammary Na⫹ and K⫹ transport; pro- take in the latter preparation operates with a Michaelis
lactin increases the mucosal-to-serosal transfer of Na⫹ constant (Km) of 40 ␮M with respect to the extracellular
across monolayers of cultured mouse mammary epithelial Pi concentration. Furthermore, Pi efflux from mammary
cells via a ouabain-sensitive pathway (24 –26). tissue can be stimulated by reversing the trans-membrane
Na⫹ gradient. These findings are consistent with the pres-
V. RELATIONS BETWEEN SODIUM, POTASSIUM, ence of a Na⫹-Pi cotransport system in the basolateral
AND CHLORIDE TRANSPORT AND WATER aspect of the mammary epithelium. Na⫹-Pi cotransport
MOVEMENTS mechanisms have been identified in a number of epithelia
including the intestine, kidney, and placenta (21, 23, 33),
It is recognized that the synthesis and secretion of several of which have been cloned and sequenced (125).
lactose is responsible for the majority of water in milk. There are, however, several differences between the
However, this cannot be the whole story, since water still properties of the Na⫹-dependent Pi transporter in mam-
appears in the milk of those species (i.e., pinnipeds) mary tissue and those described in other epithelia. First,
which contain little or even no lactose or any other sugar Pi uptake by mammary tissue is only weakly inhibited by
(151). In a number of secretory epithelia, it is accepted structural analogs; arsenate and phosphonoformic acid
that the intracellular accumulation of Cl⫺, via Na⫹-K⫹-Cl⫺ (PFA) were found to be weak or ineffective inhibitors of
cotransport across the basolateral membrane, is the driv- Pi transport. In contrast, both of these compounds have
ing force for the secretion of ions and water across the been shown to be high-affinity blockers of Na⫹-dependent
apical membrane (43, 153). The same process could occur Pi transport in other tissues (103). Second, the kinetic
in the mammary secretory cell, since the necessary gra- parameters, with respect to Na⫹ activation, are markedly
dients exist. Therefore, a situation could arise whereby different; the stimulation of Pi uptake by external Na⫹ did
the secretion of water by the mammary gland could be not exhibit sigmoidal kinetics as found, for example, in
driven partly by the secretion of lactose and partly by the the kidney (125).
secretion of ions. Variations in the concentrations of lac- Under certain experimental conditions Pi transport in
tose and ions within and between species could be related lactating rat mammary tissue occurs via an anion-ex-
to the relative flux between these two mechanisms. This change system (210). External Pi is able to trans-acceler-
hypothesis, together with the involvement of paracellular ate Pi efflux via a mechanism sensitive to DIDS. It appears
ion flow in some species, may explain how secretion of that the Pi self-exchange system is distinct from the anion
the aqueous phase is achieved in all mammals. exchanger that accepts Cl⫺ and SO2⫺ 4 as substrates (194),
since Pi efflux is not trans-stimulated by Cl⫺ and SO2⫺ 4
and Pi does not trans-accelerate SO2⫺ 4 efflux (210). More-
VI. PHOSPHATE TRANSPORT
over, the finding that the Pi self-exchanger is not Na⫹
The suckling requires a considerable amount of phos- dependent suggests that the exchange mechanism is dis-
phate for normal growth and development. There are at tinct from the Na⫹-Pi cotransport system. However, the
least three chemical forms of phosphate in milk: free physiological significance of the Pi exchanger remains to
inorganic orthophosphate (Pi) in solution as HPO2⫺ and be established.
4
H2PO⫺ The finding that Pi in milk is higher than that of
4 ; colloidal phosphate, the majority of which is cal-
cium phosphate associated with casein micelles; and ca- plasma could, at first sight, be taken as evidence that the
sein phosphate (86). For example, of the total phosphate apical membrane expresses a Pi transport mechanism.
(20.5 mM) present in goat’s milk, 6.7 mM is Pi, 8.9 mM is However, the major pathway for Pi secretion into milk is
colloidal, and 4.9 mM is casein bound (146). Despite the believed to be via the Golgi vesicle route. This hypothesis
complex nature of phosphate in milk, it can be predicted is based on two important findings. First, Neville and
that milk phosphate is derived from plasma Pi that has to Peaker (146) found that the apical aspect of the lactating
be transported across the basolateral membranes of the goat mammary epithelium is almost impermeable to Pi.
secretory cells. In accordance with this prediction, the Second, the time course of Pi secretion into milk, follow-
mammary gland has the capacity to extract large quanti- ing an intravascular injection of radiolabeled Pi, was sim-
ties of Pi from blood. For example, it is estimated that the ilar to that of casein phosphate and colloidal phosphate.
entire pool of plasma Pi is replaced every 10 min in This finding raises questions about the nature of Pi trans-
lactating rats (30). port across the Golgi membrane. There is, of course, the
The mechanism of Pi transport by the lactating mam- possibility that Golgi secretory vesicles do not need to
mary gland has only recently received attention. Pi uptake transport Pi per se from the cytosol because sufficient Pi
by rat mammary tissue explants and the perfused rat is generated in the Golgi lumen by the hydrolysis of UDP
mammary gland occurs predominantly via a Na⫹-depen- during lactose synthesis (Fig. 4). However, this mode of Pi
dent process (210).The Na⫹-dependent moiety of Pi up- generation will not occur in the Golgi lumen of pinnipeds.

calcium present in milk (144). Neville and Peaker (146)

found that the mammary gland does not transport calcium
in the milk-to-blood direction, suggesting that calcium
cannot cross the apical membrane or tight junctions. Thus
it can be inferred that calcium transport into milk is
transcellular. Furthermore, because the time course of
appearance of labeled milk calcium is similar to that of
casein and lactose, it has been suggested that calcium is
secreted via the Golgi vesicle route (146). It follows that
casein-bound calcium and colloidal calcium, along with
free ionized calcium, are all derived from plasma Ca2⫹,
indicating that Ca2⫹ must be able to cross both the baso-
lateral and Golgi membranes before secretion.
The first question that arises is, What mechanism
transports calcium across the basolateral membrane?
Surprisingly, there is a paucity of information about the
system(s) available for calcium transport across the
blood-facing membrane except for the finding that it is a
temperature-sensitive process that increases during the
transition from pregnancy to lactation (147). Whatever
the nature of the system(s), it must operate with a high
capacity given the large amount of calcium secreted into
milk. Calcium channels would of course fulfill such a role.
However, until the mechanism of calcium uptake by mam-
mary secretory cells is thoroughly studied, the presence
of calcium channels in the basolateral membrane can only
remain a suggestion.
Mammary epithelial cells maintain intracellular Ca2⫹
at a low concentration (56, 75). This observation prompts
the next question, How do mammary epithelial cells main-
tain a low cytosolic Ca2⫹ concentration while facilitating
FIG. 4. A schematic diagram of the mechanisms that may be in- a large transcellular calcium flux? A low calcium concen-
volved in the transport and metabolism of phosphate and Ca2⫹ in
mammary secretory cells. Phosphate is transported across the basolat-
tration could be maintained if Ca2⫹ were pumped into
eral membrane via a (Na⫹-Pi) cotransport mechanism and possibly by an intracellular organelles at a rate that matched the entry of
anion-exchange system. Pi is formed within the lumen of the Golgi calcium across the basolateral membranes of the secre-
during the synthesis of lactose as a consequence of the hydrolysis of
UDP-galactose. In addition, Pi is generated during the process of casein
tory cells. In this connection, there is evidence suggesting
phosphorylation. The mechanism of Ca2⫹ uptake across the basolateral that Golgi membranes express an ATP-driven Ca2⫹ pump
membrane has not been precisely identified but may be a Ca2⫹ channel. (149, 241, 245). Golgi-derived vesicles are able to accumu-
Stretch-activated Ca2⫹ channels may also provide a pathway for Ca2⫹
uptake. Ca2⫹ is pumped across Golgi membranes by a Ca2⫹-ATPase. Pi
late Ca2⫹ by a saturable process (Km ⫽ 0.8 ␮M) that
and Ca2⫹ are transported into milk along with lactose, casein, and requires the hydrolysis of ATP (Fig. 4). A Ca2⫹ pump
citrate after exocytosis of Golgi-derived vesicles at the apical aspect of situated in the basolateral membranes of mammary secre-
the secretory cells. LS, lactose synthase; CK, casein kinase; A, ADPase.
tory cells could also act to keep intracellular Ca2⫹ at a low
concentration. However, no evidence for such a transport
VII. CALCIUM TRANSPORT system has been presented. In addition, no evidence for a
basolateral Na⫹/Ca2⫹ exchanger, another transport sys-
Milk is a calcium-rich fluid (94). For example, the tem which could regulate intracellular Ca2⫹, has been
total calcium concentration in rabbit’s milk can approach found (143).
a concentration of 100 mM. Accordingly, during lactation, Ca2⫹ uptake by mammary tissue may be a hormon-
the mammary gland extracts large quantities of calcium ally regulated process. Ca2⫹ accumulation by cultured
from plasma to meet the requirements of the growing mouse mammary explants, presumably via the basolateral
neonate. Milk calcium, like phosphate, exists in several membranes, is stimulated by 1,25-(OH)2D3, the hormonal
chemical forms: free ionized calcium, casein-bound cal- form of vitamin D, in a dose-dependent fashion (137), and
cium, and calcium complexed to inorganic anions such as Ca2⫹ transfer into goat’s milk is increased by parathyroid
phosphate and citrate. In most milks, calcium associated hormone-related protein (15). However, in both of these
with casein micelles comprises the majority of the total examples, the target mechanism is unknown.

VIII. CITRATE TRANSPORT X. STRETCH-SENSITIVE ION TRANSPORT

MECHANISMS
Citrate exists in the aqueous phase of milk in a
variety of chemical forms such as calcium citrate⫺, ci- Milk accumulation in the alveolar lumen leads to
trate2⫺, and citrate3⫺. The relative proportion of each distension of the secretory alveoli, and cellular deforma-
chemical form depends on factors such as H⫹, Ca2⫹, and tion may play an important role in determining secretory
Mg2⫹ and thus citrate can be considered as an important activity during the cessation of lactation. An increase in
milk buffer. The lactating goat mammary epithelium is intramammary pressure at the end of lactation has been
impermeable to citrate in both directions, suggesting that shown to reduce the rate of milk secretion in the goat
citrate is synthesized within the secretory cells and re- (161), suggesting that, in addition to local, autocrine
leased into milk after exocytosis of Golgi vesicles (116). chemical control of milk secretion by the milk protein
In this connection, a Golgi-enriched fraction isolated from FIL, mammary distension could also play a part in con-
the bovine mammary gland accumulated citrate in a fash- trolling the rate of milk secretion in response to demand
ion sensitive to temperature, suggesting that a citrate by the young (48, 163, 168, 169, 247–249).
transport system may be present in the Golgi apparatus Cell distension has marked effects on mammary
membrane (267). The putative citrate transport mecha- gland ion transport. Enomoto et al. (58) found that me-
nism in the Golgi membranes remains to be fully charac- chanical stimulation of cultured mouse mammary cells,
terized. induced by touching with a blunt microelectrode, resulted
in a short depolarization (1– 8 s) followed by a prolonged
(10 – 40 s) hyperpolarization. At present, the locus of the
IX. IODIDE TRANSPORT stretch-sensitive channels (apical and/or basolateral) on
the mammary epithelial cells is not known. The mecha-
The concentration of iodide in milk can be 20- to nism underlying the depolarization remains to be deter-
30-fold higher than that found in maternal plasma (32), mined, but it has been established that the hyperpolariza-
matching the requirements of the suckling for the synthe- tion can be attributed to the activation of quinine-
sis of thyroid hormones. Iodide transport was investi- sensitive, Ca2⫹-dependent K⫹ channels (58, 59, 72).
gated relatively early because of the contamination of Accordingly, mechanical stimulation of cultured mouse
land and milk supplies by radioactive iodine isotopes mammary cells leads to an increase in the intracellular
from nuclear explosions or accidents. In addition, it was Ca2⫹ concentration. However, the increase in the Ca2⫹
thought that an understanding of the mammary iodide concentration is not confined to the touched cell but also
transport mechanism could help limit the transfer of ra- spreads to surrounding cells, even to those that have no
diolabeled iodide into the food chain via milk after envi- direct physical contact with the stimulated cell, suggest-
ronmental contamination. ing that a paracrine factor may be involved (60, 73). There
The iodide transport system in the thyroid gland has is evidence that ATP is released from cultured mammary
recently been cloned (47). It is a Na⫹-dependent mecha- cells upon mechanical stimulation (73). In this connec-
nism sensitive to anions such as perchlorate and thiocy- tion, extracellular ATP, acting via P2-purinergic receptors,
anate (155). A body of evidence suggests that mammary has been shown to increase intracellular Ca2⫹ in human
epithelial cells express a similar iodide transport mecha- breast tumor cells (70). Cell swelling, induced by hypos-
nism. The lactating mammary gland has the ability to motic shock, increases the intracellular Ca2⫹ concentra-
accumulate iodide into milk against a high concentration tion in acini isolated from lactating mouse mammary
gradient by a system that is inhibited by perchlorate and gland (221). The effect consists of a rapid and large tran-
thiocyanate (31, 32, 53, 114). Iodide uptake by mammary sient increase in intracellular Ca2⫹ followed by a sus-
tissue explants prepared from midpregnant mice is a Na⫹- tained plateau phase. The increase in Ca2⫹ is due to
dependent process stimulated by prolactin (184, 185). increased Ca2⫹ influx from the incubation medium rather
Moreover, mRNA encoding for the Na⫹-dependent iodide than release from intracellular stores. The nature and
transporter has been identified in the lactating human precise locus of the Ca2⫹ transport pathway is unknown,
mammary gland (215). but it is insensitive to nifedipine (221). Moreover, it is not
Although the Na⫹-dependent mechanism is probably yet known if the primary stimulus is membrane stretch or
the predominant pathway for iodide uptake by mammary a change in the cellular hydration state.
cells, there is evidence that iodide also enters via a DIDS- Based on these still sparse observations, we hypoth-
sensitive anion-exchange pathway; sulfate efflux from rat esize that stretch induces ATP release into the milk or
mammary tissue can be stimulated by extracellular iodide interstitial space, where it interacts with neighboring cells
(194). leading to an increase in calcium entry into these cells.

XI. CHOLINE AND CARNITINE TRANSPORT activity of the Na⫹-dependent L-carnitine transport sys-
tem was found to decrease as lactation progressed (201).
The organic cation choline is required by the suckling L-Carnitine efflux from lactating rat mammary tissue
for normal growth and development including the synthe- is a slow process that is Na⫹ independent and cannot be
sis of ACh, phosphatidylcholine, and sphingomyelin (258). stimulated by external L-carnitine. However, cell-swelling,
The concentrations of choline (and its metabolites) in induced by a hyposmotic shock, increases the fractional
rat’s milk and plasma are 1.5 mM and 12 ␮M, respectively, release of radiolabeled L-carnitine from rat mammary tis-
suggesting that lactation may put a considerable strain on sue (201). The possibility that volume-activated L-carni-
the maternal choline stores (259). The concentration of tine release is mediated via the volume-sensitive amino
free choline in rat’s milk, 182 ␮M, is consistent with active acid transport pathway should be investigated.
choline transport. Lactating rat mammary epithelial cells
possess a high-affinity (Km ⫽ 35 ␮M), Na⫹-dependent
choline carrier that can be inhibited by hemicholinium-3 XII. GLUCOSE TRANSPORT
(41). Rat mammary epithelial cells can maintain high
concentrations of choline (41), suggesting that the Na⫹- The concentration of lactose in the milk of most
dependent carrier is situated in the basolateral membrane species is relatively high, and lactose is one of the major
and that the transepithelial choline gradient is generated milk osmolytes; for example, the concentrations in hu-
at the basal membrane of the mammary cell. man and bovine milk are, respectively, 204 and 140 mM
L-Carnitine (4-N-trimethylammonia-3-hydroxybutano-
(158). Lactose is synthesized within the lumen of the
ate) is essential for the transport of long-chain fatty acids Golgi apparatus by an enzyme complex collectively
between cell compartments. For example, the formation known as lactose synthase. Exocytosis of the Golgi secre-
of long-chain acylcarnitines from their respective coen- tory vesicles then occurs at the luminal side of the cell.
zyme A esters enables the acyl moieties to cross the Golgi and apical membranes of mammary secretory cells
mitochondrial membrane where they are subjected to are impermeable to lactose; therefore, because these
␤-oxidation (256). L-Carnitine-dependent oxidation of membranes are freely permeable to water, the volume of
long-chain fatty acids is particularly important for brain milk secreted is closely related to the rate of lactose
development in the neonate. The transport of L-carnitine synthesis.
into milk is physiologically significant because the new- The lactating mammary gland has a high demand for
D-glucose because it is the obligate precursor for lactose
born has a limited capacity for L-carnitine biosynthesis. In
rats, the lactating female depletes its reserves of L-carni- synthesis and, depending on species, is also used to syn-
tine through secretion into milk; the rate of L-carnitine thesize lipids and amino acids (110, 115). Glucose trans-
secretion is high during the first 3– 4 days of lactation but port across both the basolateral and Golgi membranes of
thereafter falls as the young develop the capacity to syn- the secretory cells is therefore an important process in
thesize L-carnitine (186). In the early stages of lactation, milk secretion.
the L-carnitine concentration in rat milk is almost an order
of magnitude greater than that of plasma (320 vs. 35 ␮M), A. Glucose Transporters
suggesting that mammary epithelial cells possess a carrier
for L-carnitine. In accordance with this prediction, a con- 1. GLUT1 expression
centrative, Na⫹-dependent L-carnitine carrier has been
identified in mammary explants isolated from rats during Lactating mammary tissue transports D-glucose via a
the early phase of lactation (201, 257). The carrier oper- facilitative transport system similar, if not identical, to
ates with a Km of 132 ␮M and maximum velocity (Vmax) of GLUT1. Amato and Loizzi (4) found that guinea pig mam-
100 pmol 䡠 h⫺1 䡠 mg intracellular water⫺1. If, as expected, mary tissue slices are capable of transporting 2-deoxyglu-
the carrier is situated on the basolateral membrane of the cose, a substrate of GLUT1 type transporters (74), via a
alveolar cells it will not be saturated with plasma L-carni- system inhibited by cytochalasin B. Rat mammary acini
tine under physiological conditions. However, the Km also transport 2-deoxyglucose and 3-O-methyl-D-glucose
value obtained with explants is probably an overestimate via a temperature-sensitive and saturable mechanism (Km
because of the presence of unstirred fluid layers. The rat for 2-deoxyglucose ⫽ 16 mM) that is inhibited by cytocha-
mammary L-carnitine carrier is not stereospecific, since lasin B and phloretin (225). Carrier-mediated D-glucose
D-carnitine is an effective cis-inhibitor. In addition, ace- uptake with characteristics of GLUT1 has also been de-
tylcarnitine, but not choline or taurine, inhibits L-carnitine scribed in mouse mammary acini (177); thus 3-O-methyl-
uptake by rat mammary explants. These characteristics D-glucose uptake by mouse mammary cells is via a satu-
suggest that the mammary tissue L-carnitine carrier is rable (Km ⫽ 14 mM), cytochalasin B-sensitive pathway.
similar to that of renal brush-border membrane vesicles, In accordance with the flux studies quoted in the
neuroblastoma cells, and JAR cells (140, 174, 182). The previous paragraph, it has been shown that rat mammary

tissue expresses mRNA that hybridizes with a cDNA (198). 1) The efflux of 3-O-methyl-D-glucose from lactating
probe for GLUT1 as well as a protein that reacts with an rat mammary tissue explants is stimulated by reversing
antibody raised against the COOH terminus region of the the transmembrane Na⫹ gradient. This finding is consis-
human erythrocyte glucose GLUT1 transporter (35, 124). tent with a Na⫹-dependent D-glucose transporter operat-
Immunofluorescence and immunoelectron studies have ing in the reverse mode or the inhibition of D-glucose
shown that GLUT1 expression is on the basolateral aspect (re)uptake via a Na⫹-dependent mechanism. 2) mRNA
of the lactating rat mammary epithelium (40, 222), con- extracted from the rat mammary gland contains a 4-kb
sistent with the finding that 2-deoxyglucose transport oc- transcript that hybridizes with the cDNA for the rabbit
curs at the blood-facing aspect of the intact mammary intestinal Na⫹-dependent D-glucose transporter (SGLT1).
gland (226). There are also several reports of GLUT1 3) A protein reacting specifically with an antibody raised
expression, both at the mRNA and protein level, in lactat- against a synthetic hydrophilic nonadecapeptide corre-
ing bovine mammary tissue (262–264). sponding to a proposed extramembranous loop of the
Although the available evidence suggests that GLUT1 rabbit intestinal SGLT1 has been found in Western blots
mediates the majority of D-glucose uptake by lactating of rat mammary cell membranes. 4) RT-PCR, using prim-
mammary tissue, this conclusion would be strengthened ers derived from the sequence of lamb intestinal SGLT1,
if sensitivity to phloretin and cytochalasin could be dem- generated a product (670 bp) from total rat mammary
onstrated in the perfused lactating mammary gland and if RNA identical in size to that obtained from intestinal RNA
the sequence of the lactating mammary tissue GLUT1 (211). Moreover, the sequence was identical to the corre-
mRNA were known. It is notable, however, that guinea pig sponding region of intestinal SGLT1. In addition, the
mammary tissue apparently has more than one type of ovine mammary gland also expresses a SGLT1 type D-
saturable transport system for D-glucose (4). Thus cy- glucose transporter; the sequence is 1,995 bp in length and
tochalasin B and phloretin do not completely inhibit the has the potential to code for a protein containing 664
saturable component of 2-deoxyglucose uptake by guinea amino acids with a sequence identical to that of the lamb
pig mammary tissue explants. In addition, a significant intestinal SGLT1 (211). SGLT1 mRNA has also been found
proportion of 2-deoxyglucose uptake by guinea pig mam- in lactating bovine mammary tissue (265).
mary tissue slices is via a nonsaturable pathway. There- The precise locus and functional significance of the
fore, it appears that GLUT1 may not be the sole mecha- mammary SGLT1-like transporter remains to be deter-
nism for glucose transport across the basolateral mined. If it is situated in the basolateral aspect of the
membrane of mammary secretory cells. mammary epithelium, then it may operate in parallel with
GLUT1 to provide mammary acinar cells with D-glucose.
2. GLUT4 expression However, an intracellular or apical location cannot be
ruled out at this stage.
No evidence has been found for the insulin-sensitive
D-glucose transporter GLUT4 in lactating rat mammary
epithelial cells (35, 40, 124). However, GLUT4 mRNA is B. D-GlucoseTransport Across the
expressed in mammary tissue taken from virgin and preg- Apical Membrane
nant rats, but expression decreases as gestation
progresses, suggesting that GLUT4 is expressed in mam- D-Glucose, along with a number of other monosac-
mary adipocytes rather than mammary epithelial cells (35, charides, is present in milk of most species at a much
40). No detectable expression of GLUT4 mRNA is evident lower concentration than that found in plasma (65). The
in lactating bovine mammary tissue (263). available experimental evidence is consistent with the
presence of a D-glucose transport system on the apical
3. GLUT2, GLUT3, and GLUT5 expression membrane of mammary secretory cells. Indeed, it has
Lactating rat mammary epithelial cells do not appear been suggested that milk D-glucose concentrations can be
to express GLUT2 or GLUT5 (35, 40). Similarly, lactating taken as a measure of the intracellular concentration (65,
bovine mammary tissue does not express GLUT2 mRNA. 142). D-Glucose and galactose introduced into the lactat-
Although low levels of GLUT3 and GLUT5 mRNA are ing goat mammary gland via the teat canal are rapidly lost
found in the bovine mammary gland, the cellular location from the milk space. The finding that fructose does not
(i.e., epithelial or stromal) of these transcripts in the gland cross the mammary epithelium, even after 16 h, suggests
is not known (263). that D-glucose and galactose cross the mammary epithe-
lium via a transcellular route and not via a paracellular
pathway (64). This conclusion is supported by the finding
4. SGLT1 expression
that radiolabeled 3-O-methyl-D-glucose introduced into
There is evidence for the presence of a Na⫹-depen- the milk space of the goat mammary gland via the teat
dent glucose transporter in lactating rat mammary tissue enters venous blood at a faster rate than radiolabeled

sorbitol (65). The nature and significance of D-glucose creased during progression from the pregnant to the mid-
transport across the apical membrane remains to be de- lactating state (177). A sharp decline in uptake was
termined. If D-glucose is able to cross the apical mem- observed in the immediate postlactational period. A
brane of rat mammary epithelial cells, then the available change in the Vmax but not the Km of 3-O-methyl-D-glucose
evidence suggests that the pathway is one other than uptake suggests that the number of transporters is af-
GLUT1 because GLUT1 protein could not be detected on fected, rather than the affinity of the carrier, by the stage
the apical membrane (40). of lactation. The expression of GLUT1 mRNA and protein
In contrast to most species (i.e., eutherian mam- fall within 24 h of litter removal in the rat (40). These
mals), the Tammar wallaby can, in late lactation, maintain observations suggest that the differences in D-glucose
higher concentrations of glucose and galactose in milk transport by mammary tissue during development, lacta-
than those in plasma (133, 134). This finding raises inter- tion, and involution reflect the expression and function of
esting questions about the nature of D-glucose and galac- GLUT1 under the prevailing hormonal milieu of these
tose transport across the apical membrane of the mam- physiological states.
mary epithelium in this species and other marsupials. For Milk secretion in the rat is controlled by both prolac-
example, why aren’t these monosaccharides rapidly lost tin and growth hormone. Fawcett et al. (66) investigated
from the milk space as would be predicted if the apical the effect of these hormones on the expression of GLUT1
membrane possessed a facilitative transport mechanism? protein in mammary secretory cell membranes isolated
from lactating rats receiving various treatments for 48 h.
Administration of bromocriptine reduced GLUT1 in mam-
C. Transport of D-Glucose Across the mary membranes, as would be expected. In contrast,
Golgi Membrane GLUT1 levels in membranes from animals that had been
treated with an anti-rat growth hormone serum were not
D-Glucose must cross the Golgi membrane to reach
significantly altered. However, when the two treatments
the site of lactose synthesis. A Golgi vesicle fraction were combined, GLUT1 expression was reduced to a level
prepared from lactating rat mammary tissue was found to below that found with bromocriptine treatment alone.
transport D-glucose; it was postulated that a pore was The results suggest that prolactin and growth hormone
involved (246). However, the precise nature of D-glucose act synergistically to maintain GLUT1 transporter expres-
transport across the Golgi membrane remains to be es- sion in rat mammary gland plasma membranes. A role for
tablished, although one report indicates that Golgi mem- prolactin in the regulation of GLUT1 expression in cul-
branes possess a protein similar to GLUT1. Therefore, it is tured mouse mammary epithelial cells has also been re-
possible that GLUT1 transports D-glucose to the site of ported (87). Prolactin stimulates 2-deoxyglucose uptake
lactose synthesis; alternatively, Golgi-derived GLUT1 pro- by mammary gland explants isolated from midpregnant
teins may be destined for the plasma membrane (124). mice (170). Although the target mechanism was not iden-
tified, it can be predicted, on the basis of experiments
D. Control of D-Glucose Transport using rats, that the prolactin effect is on GLUT1.
Zhoa et al. (264) found that administration of bovine
Given the important role of D-glucose in the process growth hormone releasing factor, but not bovine growth
of milk secretion, it is of fundamental importance that the hormone itself, increased by 21% the amount of GLUT1
factors that control the transport of D-glucose by mam- mRNA in the lactating bovine mammary gland. However,
mary epithelial cells are elucidated. However, it is surpris- the physiological significance is unclear, since the in-
ing to find that our knowledge of this important area is crease in message was not accompanied by a rise in
relatively patchy. Nevertheless, there are a number of GLUT1 protein in mammary cell membranes.
findings which suggest that D-glucose uptake by mam- The uptake of 2-deoxyglucose by the lactating rat
mary epithelial cells is influenced by the stage of mam- mammary gland in vivo was found in one study to be
mary development and lactation (and, therefore, by infer- enhanced by insulin (226). This observation is difficult to
ence, prolactational hormones like prolactin, and growth reconcile with the failure of insulin to produce acute
hormone) as well as nutritional status. stimulation of 3-O-methyl-D-glucose transport by isolated
lactating mouse mammary epithelial cells (177). If insulin
does have a direct effect on D-glucose transport by lactat-
1. Ontogeny and hormonal control
ing mammary epithelial cells, then the target must be a
Lactose synthesis and glucose uptake by the mam- transporter other than GLUT4 (see sect. XIIA). It is possi-
mary gland increase abruptly at the time of parturition ble that insulin stimulates D-glucose uptake by the rat
and decline rapidly as the gland stops secretion before mammary gland as a result of altering the magnitude of
involution (49, 69). Accordingly, the transport of 3-O- the transmembrane D-glucose gradient rather than by af-
methly-D-glucose by isolated mouse mammary cells in- fecting the transport proteins directly (34).

2. Effects of starvation of the mammary transport systems may have unique ki-
netic properties and mechanisms of control.
Overnight starvation of lactating rats reduces 2-de-
oxyglucose and 3-O-methyl-D-glucose uptake by the mam-
mary gland by ⬎90%. (226); the arteriovenous difference A. Amino Acid Transporters
in D-glucose concentration across the mammary gland is
decreased to a similar extent (156). However, refeeding 1. Na⫹-dependent transport of neutral amino acids
starved animals rapidly restores the uptake of glucose
There is good evidence for system A-like transport of
analogs and increases the arteriovenous difference. It is
neutral amino acids in lactating mouse, rat, and bovine
apparent that the effect of overnight fasting is a conse-
mammary tissue (Table 1). System A is a Na⫹-dependent
quence of a reduction of D-glucose transport via GLUT1.
transport mechanism that prefers short-chain neutral
Thus starvation decreases cytochalasin B-sensitive 3-O-
amino acids as substrates and has the distinguishing fea-
methyl-D-glucose uptake by isolated lactating mouse
ture of being able to tolerate N-methylated amino acids
mammary epithelial cells. This reduction in transport oc-
(14). Mouse and rat mammary tissue explants transport
curs as a result of a decrease in the Vmax of uptake and is
aminoisobutyric acid (AIB) via a pathway that is both Na⫹
accompanied by a decrease in the number of cytochalasin
dependent and sensitive to methylaminoisobutyric acid
B binding sites on mammary cell plasma membranes
(MeAIB), a characteristic of system A (145, 204). How-
(175). However, it has been reported that starvation does
ever, the substrate specificity of system A in rodent mam-
not alter the total GLUT1 protein content of lactating rat
mary tissue for naturally occurring amino acids remains
mammary tissue, suggesting that the reduction in D-glu-
to be established. The lactating bovine mammary gland
cose transport induced by starvation is brought about by
also appears to express system A; mammary explants
the translocation of GLUT1 carriers from the plasma
transport MeAIB in a Na⫹-dependent fashion (18).
membrane to an intracellular site (40).
The transport of amino acids via system A may, in
part, be responsible for the accumulation of certain neu-
XIII. AMINO ACID TRANSPORT tral amino acids within mammary cells with respect to
plasma (209). In contrast to mouse, rat, and bovine mam-
The lactating mammary gland has a large demand for mary tissue, it appears that the guinea pig mammary gland
amino acids to meet the requirements of milk protein does not possess system A activity. This conclusion is
synthesis. For example, a dairy cow secreting 35 l milk/ based on the lack of MeAIB-sensitive amino acid trans-
day (by no means exceptional) with a protein content of port by the perfused guinea pig mammary gland (131).
3.3% needs over 1 kg/day of amino acids to sustain milk Bovine mammary tissue transports AIB via a system
protein synthesis. The first studies on amino acid trans- that is Na⫹ dependent but not inhibited by MeAIB (18).
port by the lactating mammary gland focused on arterio- The simplest interpretation of this experimental observa-
venous differences in amino acid concentrations and tion is that bovine mammary tissue possesses system
combining these with measurement of mammary blood ASC-like activity. Similarly, it has been concluded that
flow to establish whether amino acids were the blood lactating guinea pig mammary tissue expresses system
source of milk proteins and then whether their quantita- ASC-like activity (131). The presence of system ASC in
tive uptake was sufficient to account for milk output of mouse mammary tissue is a matter of controversy. On the
protein (129). Many studies in goats, cows, sheep, pigs, one hand, Verma and Kansal (231) have shown that me-
guinea pigs, and rats have shown that the arteriovenous thionine may utilize system ASC; on the other hand, Nev-
differences across the gland are substantial, especially for ille et al. (145) found no evidence for AIB transport via
some of the essential amino acids (82, 119, 129, 136, 240). system ASC in mouse mammary tissue. The difference in
This extraction of amino acids from blood is generated by results is difficult to reconcile but may reflect metabolism
amino acid transport across the basolateral membranes of of methionine in the former study. In this connection, no
the secretory cells. Despite the importance of amino acids evidence for system ASC in rat mammary tissue has been
to milk secretion and mammary metabolism, it was some found (204).
time before the cellular mechanisms responsible for L-Glutamine transport by the lactating rat mammary
amino acid uptake by the mammary gland received the gland is partly dependent on the presence of Na⫹ (39).
attention they deserved. Although our current under- The Na⫹-dependent component, a low-affinity pathway
standing of mammary amino acid transport systems and that has been localized to the basolateral surface of the
their regulation lags behind what is known in other tis- epithelium, is not mediated by system A, since L-glutamine
sues, it is now becoming clear that the lactating mammary transport is insensitive to MeAIB. The Na⫹-dependent
gland possesses amino acid transport systems analogous moiety of L-glutamine transport remains to be identified,
to those described in other organs such as the intestine, but a mechanism akin to system N (104) must be a strong
kidney, and placenta (209). However, it appears that some candidate.

TABLE 1. Kinetic properties of mammary tissue amino acid transport systems

Experimental Reference
Amino Acid Transport System Species Preparation Km Vmax No.
Na⫹-dependent L-glutamate uptake Rat Explants 112.5 ␮M 71.3 nmol 䡠 min⫺1 䡠 g cells⫺1 138
⫺
(system XAG )
Na⫹-dependent L-glutamate uptake Rat Perfused gland 18.1 ␮M 40.3 nmol 䡠 min ⫺1
䡠 g tissue ⫺1
138
⫺
(system XAG )
Na⫹-dependent D-aspartate uptake Rat Explants 32.4 ␮M 24.5 nmol 䡠 min⫺1 䡠 g cells⫺1 139
⫺
(system XAG )
Na⫹ ⫹ Cl⫺-dependent taurine uptake Rat Explants 37.5 ␮M 2.5 nmol 䡠 min ⫺1
䡠 g ICW ⫺1
205
(system ␤)
Na⫹-dependent taurine uptake (possibly Gerbil Explants 70 ␮M 7.9 nmol 䡠 min⫺1 䡠 g ICW⫺1 197
system ␤)
Na⫹-dependent AIB uptake (system A) Mouse Explants 2.0 mM ⫺1
197 nmol 䡠 min 䡠 g cells ⫺1
145
Na⫹-independent AIB uptake (system L) Mouse Explants 2.7 mM 150 nmol 䡠 min⫺1 䡠 g cells⫺1 145
Na⫹-dependent methionine uptake Mouse Explants 0.47 mM 18.8 nmol 䡠 min⫺1 䡠 g cells⫺1 231
(system A)
Na⫹-independent, MeAIB-sensitive Mouse Explants 0.46 mM 30.0 nmol 䡠 min⫺1 䡠 g cells⫺1 231
methionine uptake (system L)
Na⫹-dependent methionine uptake Mouse Explants 0.46 mM 12.4 nmol 䡠 min⫺1 䡠 g cells⫺1 231
(system ASC)
Na⫹-dependent tyrosine uptake (system Mouse Explants 1.67 mM 75.1 nmol 䡠 min ⫺1
䡠 g cells⫺1
100
unknown)
Na⫹-independent, BCH-insensitive Mouse Explants 15.75 mM 157.5 nmol 䡠 min⫺1 䡠 g cells⫺1 100
tyrosine uptake (system T)
Na⫹-independent, BCH-sensitive tyrosine Mouse Explants 0.23 mM 31.0 nmol 䡠 min ⫺1
䡠 g cells⫺1
100
uptake (system L)
AIB uptake (in presence of prolactin) Mouse Cultured explants 0.75 mM 430 nmol 䡠 min⫺1 䡠 g ICW⫺1 183
Km, Michaelis constant; Vmax, maximum velocity; AIB, aminoisobutyric acid; MeAIB, methylaminoisobutyric acid; BCH, 2-aminobicyclo-[2,2,1]-
heptane-2-carboxylic acid.
2. Na⫹-dependent transport of anionic amino acids molecular level may reveal that there is more than one
system (although their kinetics will be similar). However,
In some species, L-glutamate is the most abundant if it transpires that the Na⫹-dependent moiety of L-gluta-
amino acid in milk protein. In addition, the mammary mate transport in rat mammary tissue is mediated by a
gland of some species, notably humans, is able to gener-
single mechanism, then the mammary system has a phar-
ate a large transepithelial gradient of free glutamate in
macological profile unlike that of the high-affinity, Na⫹-
favor of milk (5). In vivo, the mammary gland extracts
dependent systems that have been cloned (EAAC1,
large quantities of glutamate from arterial blood (115),
GLAST, and GLT1) and characterized (97, 171, 219). The
indicating mammary cells do not rely on intracellular
evidence for such a difference is based on the effects of
L-glutamate synthesis. In lactating mammary tissue, there
dihydrokainate (DHK) and ␣-aminoadipate (AAD): an-
is convincing evidence that L-glutamate and L-aspartate
are transported by a system that is analogous to system ionic amino acid transport by mammary tissue is inhibited
⫺
XAG (138, 139); the mechanism is of high affinity, is Na⫹ by DHK but not AAD (139); EAAC1 is inhibited by AAD
dependent, and interacts only with anionic amino acids. but not DHK; GLT1 is sensitive to both AAD and DHK; and
Another XAG ⫺
-like feature of the transport system in the rat GLAST is inhibited by neither AAD nor DHK (98, 109).
is that the carrier differentiates between the optical iso- Another piece of evidence which suggests that the mam-
mers of glutamate but not those of aspartate. mary system is different from EAAC1 is the effect of
⫺ extracellular amino acids on the efflux of substrates via
XAG -like transport is the predominant, if not only,
mechanism for anionic amino acid transport in rat mam- the Na⫹-dependent carrier. L-Cysteine is capable of trans-
mary tissue; no evidence for L-glutamate transport by accelerating L-glutamate efflux via EAAT3 (260), a human
system X⫺ ⫹
c , a Na -independent transporter which cata-
homolog of EAAC1, whereas it has little or no effect on
lyzes the exchange of L-glutamate and L-cystine (11), was amino acid efflux from rat mammary tissue (139). The
found (139). Studies using the perfused lactating rat mam- mammary gland L-glutamate transport mechanism does
mary gland suggest that the system XAG ⫺
-like activity is display Na⫹-dependent exchange activity; anionic amino
situated, as predicted, in the basolateral membrane of the acids such as L-glutamate, L-aspartate, D-aspartate, and
mammary epithelium (138, 139). Although the kinetics L-cysteine sulfinate are transported by the carrier operat-
suggest that there is only a single saturable system, a ing as an exchanger (139) (Table 2). This mode of oper-
detailed study of anionic amino acid transport at the ation may allow the uptake of L-glutamate to regulate the

TABLE 2. Effect of external amino acids (at 500 ␮M) on acids including ␤-alanine and hypotaurine (205). Another
D-aspartate efflux from lactating rat mammary tissue pathway for taurine uptake in rat mammary tissue may
exist, since a small portion of the Na⫹-dependent compo-
Efflux Rate Constants, min⫺1 ⫻ 10⫺4
nent is not Cl⫺ dependent (205). The mammary taurine
Amino Acid n Control ⫹ Test Difference P Value transport system has similar characteristics to the Na⫹-
and Cl⫺-dependent taurine transporters that have been
L-Glutamate 7 75 ⫾ 10 362 ⫾ 32 287 ⫾ 38 ⬍0.001
D-Aspartate 6 82 ⫾ 12 311 ⫾ 32 229 ⫾ 43 ⬍0.01
described (and cloned) in other epithelia (122, 178, 229).
L-Aspartate 4 86 ⫾ 8 415 ⫾ 32 329 ⫾ 36 ⬍0.01 However, the molecular identity of the rat mammary tau-
CSA 4 58 ⫾ 18 369 ⫾ 55 311 ⫾ 70 ⬍0.05 rine transport mechanism(s) remains to be determined.
L-Cysteine 5 103 ⫾ 9 161 ⫾ 14 58 ⫾ 9 ⬍0.01
L-Leucine 3 80 ⫾ 12 88 ⫾ 23 8 ⫾ 11 NS
Maintenance of a high intracellular concentration of tau-
D-Glutamate 5 87 ⫾ 20 105 ⫾ 19 18 ⫾ 9 NS rine by the Na⫹-dependent system may be important in
DHK 4 54 ⫾ 11 100 ⫾ 14 46 ⫾ 3 ⬍0.001 relation to mammary cell volume regulation; there is good
AAD 3 73 ⫾ 16 87 ⫾ 22 14 ⫾ 9 NS
evidence that taurine (and other amino acids) may play a
D-Aspartate efflux was measured into a medium containing (in role following cell swelling (see XIIID).
mM) 135 NaCl, 5 KCl, 2 CaCl2, 1 MgSO4, 10 glucose, and 20 Tris-MOPS, In view of the finding that gerbil milk contains a high
pH 7.4, and then to a similar medium supplemented with a “test” amino concentration of taurine (⬃5 mM) (181), the uptake of tau-
acid at 500 ␮M. CSA, L-cysteinesulfinate; DHK, dihydrokainate; AAD,
aminoadipate. Only those amino acids that inhibited L-Glu and D-Asp rine by lactating mammary tissue from the gerbil has also
uptake, with the exception of DHK, were effective at trans-stimulating been investigated (197). Taurine uptake by gerbil mammary
⫹
D-Asp efflux, suggesting that the high-affinity Na -dependent carrier is
tissue explants, like that of the rat, is mediated by a high-
able to act as an exchanger as well as a cotransport system. The finding
that DHK did not markedly stimulate D-Asp efflux indicates that DHK is affinity (Km⫽ 70 ␮M), Na⫹-dependent transport system that
a nontransported inhibitor. [Data from Millar et al. (139).] is selective for ␤-amino acids. However, the Na⫹-dependent
moiety of taurine uptake is not Cl⫺ dependent, and anions
such as NO⫺ ⫺ ⫺
3 and SCN can substitute for Cl (197). In this
⫺ ⫹
intracellular concentration of L-aspartate (and vice versa); connection, a Cl -independent, Na -dependent taurine
however, it is not yet known if the exchange of intracel- transport system has been found in cultured human kidney
lular amino acids is required for amino acid uptake by the cells (95); however, this mechanism has a low affinity (Km⫽
high-affinity anionic amino acid carrier. 1 mM) for taurine. Similarly, a Na⫹-dependent taurine trans-
port system that is not coupled to Cl⫺ has been identified in
3. Na⫹-dependent transport of ␤-amino acids rabbit proximal kidney tubule (29).
Taurine (ethanesulfonic acid), a nonprotein ␤-amino 4. Na⫹-independent transport of neutral amino acids
acid, is present in the milk of a large number of species
(181); in some species, notably pinnipeds, exceptionally System L, a well-studied system for neutral amino
high concentrations are found, 13 mM in the Antarctic fur acids that is sensitive to 2-aminobicyclo-[2,2,1]-heptane-2-
seal for example (192). It has been suggested that the high carboxylic acid (BCH), has been identified in lactating
taurine levels in the milk of certain marine mammals are mouse, guinea pig, bovine, and rat mammary tissue (18,
related to their high milk fat content (152), since taurine 131, 145, 206, 231) (Table 1). This conclusion is based on
is required for bile salt synthesis. There is a strong pos- the finding that mammary tissue from these species ex-
sibility that milk taurine may play an important role in hibits BCH-sensitive, Na⫹-independent transport of neu-
growth and development of the suckling (90), particularly tral amino acids. System L in mammary tissue appears, on
in those species whose neonates are not capable of syn- the basis of cis-inhibition studies, to have a wide sub-
thesizing taurine. However, taurine is also present in the strate specificity and a low strereospecificity (145, 206).
milk of species whose young are able to synthesize tau- This mechanism may be the single most important path-
rine, suggesting that the transfer of the ␤-amino acid into way for the uptake of the essential neutral amino acids by
milk has other roles. the lactating gland. Studies using the perfused lactating
Lactating rat mammary tissue accumulates taurine to guinea pig and rat mammary gland preparations have
higher concentrations than plasma, generates a transepi- shown that system L is situated in the blood-facing aspect
thelial concentration gradient with milk concentrations of the mammary epithelium (39, 131). There is evidence to
higher than plasma, and transports radiolabeled taurine suggest that in bovine mammary tissue system L acts as
from blood to milk (205, 218, 220). Collectively, these an exchanger: intracellular cycloleucine is able to trans-
results suggest that the rat mammary gland possesses an stimulate the uptake of cycloleucine by bovine mammary
active transport mechanism for taurine. The predominant explants. Similarly, L-methionine uptake by mouse mam-
pathway for taurine uptake by rat mammary tissue ex- mary explants via system L is trans-accelerated by intra-
plants is a high-affinity (Km ⫽ 43 ␮M), Na⫹- and Cl⫺- cellular L-methionine (99). However, AIB efflux from rat
dependent transport mechanism selective for ␤-amino mammary tissue is not trans-stimulated by substrates of

system L (206). Likewise, L-phenylalanine egress from rat but it is evident, on the basis of cis-inhibition studies, that
mammary tissue, in the absence of extracellular Na⫹, is neutral amino acids also interact with the L-lysine carrier
not significantly affected by supplementing the incubation (208). Thus a wide range of neutral amino acids are able
medium with system L substrates (D. B. Shennan, unpub- to reduce L-lysine uptake by mammary tissue explants.
lished data). If it is assumed that system L in rat mammary The finding that L-lysine is trans-stimulated by L-leucine
tissue is identical to system L in bovine and mouse mam- and L-glutamine (even in the absence of extracellular
mary glands, then the finding that amino acid uptake, but Na⫹) suggests that neutral amino acids are actually trans-
not efflux, is trans-stimulated suggests that system L in ported by the cationic amino acid carrier rather than
mammary tissue operates with asymmetrical kinetics. simply acting as inhibitors. It is interesting to note that
This form of asymmetry between the internal and external Neville et al. (145) found that L-arginine inhibited AIB
face of the carrier favors amino acid retention by the uptake by lactating mouse mammary tissue. The cationic
mammary epithelium particularly after a postprandial in- amino acid carrier that interacts with neutral amino acids
crease in plasma amino acids. In this connection, a sulfate has also been found using the perfused rat mammary
carrier with asymmetrical kinetics for trans-stimulation gland, suggesting that it is located in the basolateral mem-
between its internal and external face has been described brane of the secretory cells (38). The available evidence
in human placental brush-border membrane vesicles (36). suggests that the cationic amino acid transport system
However, there is a distinct possibility that system L in rat that interacts with neutral amino acids in lactating rat
mammary tissue may not be able to act as an exchange mammary tissue is not system y⫹. However, the uptake of
mechanism, suggesting that there may be tissue-specific cationic amino acids by rat mammary tissue may be me-
variants of the transport mechanism. This is already ap- diated by more than one pathway as the kinetics of L-
parent with respect to substrate specificity given that lysine transport have not been thoroughly examined. The
system L in some tissues (e.g., the human placenta) does molecular identity of the rat mammary tissue cationic
not transport AIB (57). amino acid transport mechanism remains to be identified,
System T, a Na⫹-independent mechanism originally but there is the possibility that the system may be related
identified in human erythrocytes (189), has been de- to one of the amino acid transport systems (bo⫹, y⫹L) that
scribed in mouse mammary tissue (100). Thus the portion accepts both cationic and neutral amino acids as sub-
of L-tyrosine uptake by mouse mammary tissue explants strates (22, 52). On a cautionary note, it must be borne in
that is not dependent on extracellular Na⫹ and is not mind that metabolism of L-lysine (particularly in the ex-
inhibited by BCH has been ascribed to system T. This periments employing explants) may have confounded the
mechanism prefers aromatic amino acids (L-tyrosine, L- interpretation of the results.
tryptophan, and L-phenylalanine) as substrates.
B. Amino Acid Transport Across the Apical
5. Cationic amino acid transport Membrane of Mammary Secretory Cells
The transport of cationic amino acids by mammary To date, the amino acid transport systems that have
tissue is of particular interest because it has been shown been identified in mammary tissue probably reside in the
in several species that L-arginine is taken up by mammary blood-facing aspect of the mammary epithelium. How-
tissue in excess of its requirement for milk protein syn- ever, the fact remains that the milk of some species
thesis (45). It has been suggested that L-arginine is used as contains significant quantities of free amino acids. A strik-
a precursor for amino acids whose uptake is less than that ing example is the milk of pinnipeds, where the total
required for milk protein synthesis as well as for urea concentration of free amino acids is in the range 12–21
(115, 130). In addition, L-arginine is needed by mammary mM (192). Invariably, the most abundant free amino acids
epithelial cells for the synthesis of nitric oxide (112). The in milk are taurine and L-glutamate (181, 192). Although
transport of cationic amino acids by lactating bovine and paired measurements of free amino acids in milk and
rat mammary tissue does not depend on Na⫹ (17, 208). plasma from single animals are lacking, it would appear
L-Lysine and L-arginine (and ornithine) share a pathway that some amino acids are concentrated in milk. For
for transport in bovine mammary tissue that is not cis- example, L-glutamate is concentrated in human milk (5).
inhibited by MeAIB and L-histidine (but possibly by c-Leu) There is the possibility that amino acids can cross the
(17). However, the results of these experiments must be apical membrane via specific carriers, but no studies have
interpreted with caution, since the effects of the amino been reported.
acids were not tested on the initial rate of L-lysine uptake.
Nevertheless, Baumrucker (17) concluded that the cat- C. Control of Amino Acid Transport
ionic amino acid transport in bovine mammary tissue is
mediated via system y⫹. L-Lysine and L-arginine also share Amino acid uptake by lactating mammary tissue is
a pathway for transport in lactating rat mammary tissue, regulated by a variety of factors including prolactin, milk

stasis, and starvation. Most of the studies have involved D. Volume-Sensitive Amino Acid Transport
measurement of arteriovenous differences, sometimes
combined with blood flow determinations, and it was not Most cells regulate their volume after swelling or
possible to identify which transport systems were af- shrinking. Perturbations induced by anisosmotic condi-
fected. Nevertheless, it is possible in a few cases to be tions and substrate accumulation activate membrane
almost certain what pathways would have been involved; transport mechanisms (85, 106). Swelling activates the
for example, if a manipulation affected the extraction of release of K⫹, Cl⫺, and organic osmolytes such as amino
anionic amino acids, then it can safely be assumed that acids (especially taurine); this solute efflux together with
the effect was on the high-affinity, Na⫹-dependent, an- osmotically obliged water returns cells to their normal
ionic amino acid transporter given that this is the only volume. Shrinkage has the opposite effect, leading to the
pathway available for this class of amino acids. uptake of solutes such as Na⫹, K⫹, Cl⫺, and amino acids
Because prolactin is of major importance to milk and, consequently, water.
secretion in rats, it is not surprising that full secretory Mammary tissue has a relatively high intracellular
activity stimulated by prolactin is associated with free amino acid pool (93, 207, 209). In particular, the
changes to amino acid transport. Treating rats with bro- concentrations of nonessential amino acids are higher
mocriptine during peak lactation, to block prolactin re- than in plasma or milk. It appears that mammary tissue
lease, reduced the arteriovenous concentration difference may utilize amino acids to regulate volume after swelling
across the mammary gland for all amino acids with the because exposing rat mammary explants to hyposmotic
exception of L-aspartate, L-glutamate, and L-valine. How- conditions increased the efflux of radiolabeled taurine
ever, the arteriovenous differences were restored (but not and glycine in a rapid and reversible manner (207). This
for L-threonine, L-isoleucine, and L-leucine) by exogenous efflux is seen in the perfused rat mammary gland, sug-
prolactin (234). AIB uptake by mammary tissue explants gesting that the amino acids were exiting via the basolat-
from rats treated with bromocriptine was reduced; the eral membrane of the secretory cell (39). However, the
magnitude of the contribution of amino acids to mam-
reduction in AIB uptake is attributable to amino acid
mary cell volume regulation is not known.
transport systems A and L (206). Similarly, c-Leu trans-
It is not yet known if the amino acids whose transport
port by cultured rat mammary acini isolated from lactat-
is affected by cell swelling utilize one or more pathways. The
ing rats during late lactation can be stimulated by prolac-
simplest explanation of the available evidence is that amino
tin, a finding consistent with an increase in system L
acids, such as taurine and glycine, exit mammary tissue via
(187). Amino acid transport by mammary tissue taken
a single pathway. Volume-sensitive taurine efflux from mam-
from midpregnant mice also responds to prolactin; AIB
mary tissue may be channel mediated, since the process is
transport was increased but could be inhibited by cyclo-
relatively temperature insensitive and does not exhibit
heximide and actinomycin D (183). trans-stimulation (207). However, the exact identity of the
As would be expected from the arrest of secretion mammary tissue pathway is unknown. Apparently, volume-
that occurs in the glands of rats that are sealed to prevent activated taurine efflux from some, but not all, cell types is
milk removal, the uptake (as interpreted from arterio- via volume-activated anion channels. For example, it is be-
venous differences) of all amino acids was reduced (240). yond doubt that taurine is able to traverse anion channels in
Similarly, starvation for 24 h decreased the arteriovenous Madin-Darby canine kidney, C6 glioma, and inner medullary
difference in concentration of all the amino acids in a collecting duct cells (9, 28, 92). The finding that volume-
reversible manner (237). Consistent with the in vivo ob- activated taurine efflux is also inhibited by anion transport
servations, it was subsequently shown that the uptake of blockers such as DIDS, 5-nitro-2(3-phenylpropylamino) ben-
radiolabeled AIB by mammary acini isolated from starved zoic acid (NPPB), and niflumic acid has also been used to
lactating rats is markedly reduced (223, 239). It has been support the claim that amino acids use volume-activated
suggested that the reduction in amino acid transport can anion channels (81, 107, 108, 157, 191). However, care must
be attributed to an increase in the circulating concentra- be exercised in interpreting studies employing anion trans-
tions of ketone bodies (237). In contrast, in the mouse, port inhibitors because these compounds are notoriously
Verma and Kansal (232) found that L-methionine uptake nonspecific (37). Interestingly, volume-sensitive taurine ef-
by mammary explants from animals that had been flux from lactating rat mammary is inhibited by DIDS, NPPB,
starved, for the very long period of 48 h, was markedly and niflumic acid, albeit at relatively high concentrations.
stimulated. It appeared that the increase in uptake can be This could, at first sight, be taken as evidence that volume-
attributed to systems A and ASC (but not system L) and sensitive taurine efflux from mammary tissue utilizes anion
reflects an increase in the Vmax of the transport mecha- channels. However, Shennan and co-workers (200, 207)
nisms. These results are difficult to reconcile with those found that the effluxes of radiolabeled I⫺ and D-aspartate,
found using the rat but could be associated with the substrates of volume-activated anion channels, were not
longer period of starvation. increased when mammary explants were subjected to a

been suggested that amino acids derived from circulating

peptides (and/or proteins) may make up the apparent
shortfall. There is good evidence that amino acids derived
from intravenously administered dipeptides can be incor-
porated into milk proteins in the goat (7, 8). The possible
mechanisms for this incorporation are transport of intact
peptides with intracellular hydrolysis and extracellular
hydrolysis followed by uptake of the free amino acids. In
this connection, Wang et al. (243) claim to have evidence,
albeit indirect, for the transport of intact peptides by
mouse mammary tissue explants; however, the mecha-
nism(s) remains to be characterized.
In a recent study, an attempt was made to quantify
the transport of intact peptides across the basolateral
membrane of rat mammary secretory cells (199). The
transport of dipeptides containing a D-isomer at the NH2
terminus, a configuration which confers resistance to hy-
FIG. 5. Effect of a hyposmotic shock (307–182 mosmol/kgH2O) on drolysis (132), by the perfused lactating rat mammary
the fractional release of radiolabeled taurine (Œ) and D-aspartate (F)
from the perfused lactating rat mammary gland. Data are means ⫾ SE
gland was examined. This experimental approach demon-
(n ⫽ 3). The results suggest that the volume-activated system is selective strated dipeptide transport by a low-affinity pathway.
for taurine and is situated in the basolateral aspect of the mammary However, the rate of peptide transport was low compared
epithelium. [From Calvert and Shennan (39).]
with the uptake of free amino acids (199). The results
suggest that the pathway for dipeptide transport across
the basolateral membrane of rat mammary secretory cells
hyposmotic challenge, and it was concluded that volume-
is different from the peptide transport systems that have
sensitive amino acid efflux from mammary tissue does not
been cloned from the intestine (PepT1) and kidney
utilize anion channels (Fig. 5). Volume-activated amino acid
(PepT2) (67, 123).
efflux independent of anion channels has been demon-
The study by Shennan et al. (199) does not rule out a
strated in other cell types (51, 113, 188). It must be borne in
role for a peptide transport system such as PepT1 or
mind that the experiments of Shennan and co-workers (207,
PepT2 in the apical membrane of mammary secretory
209) do not rule out the existence of volume-activated anion
cells. A specific peptide transporter on the apical aspect
channels in rat mammary tissue, only that amino acids do
of the epithelium could act as a scavenger system to
not appear to use that route. In this connection, Kulpa (111)
transport the products of milk protein hydrolysis back
has recently shown using the whole cell, patch-clamp re-
into secretory cells. It is interesting to note that a favor-
cording technique that cultured bovine mammary cells pos-
able pH gradient exists across the apical membrane for
sess volume-sensitive Cl⫺ channels.
proton-coupled peptide transport, and rabbit mammary
gland has been reported to express mRNA encoding the
E. Peptide Transport high-affinity PepT2 transport system (54). However, the
location of the transporter remains to be established.
Because of the nature of the digestive system and the It has been suggested that peptides may cross the ba-
diverse sources of protein available to ruminant animals, solateral membranes of epithelia via an anion transporter
particular diets can be devised that alter the availability of (132). Support for this hypothesis comes from the finding
amino acids. Dairy animals have very large mammary that a range of aromatic peptides and cephalosporin antibi-
glands for their body size (83) and, therefore, extract otics are able to interact with the renal organic anion trans-
considerable quantities of amino acids from blood. Early porter (230). In this connection, benzylpenicillin, an antibi-
studies in the goat and cow showed that “the mammary otic which contains an amide bond, is actively transported
uptake of essential amino acids is sufficient to account for from blood to milk across the lactating caprine mammary
the output of their corresponding residues in the protein” gland via a pathway sensitive to probenecid, an inhibitor of
(129). Later, there have been suggestions that the mam- the renal organic anion transport system (193). Similarly,
mary uptake of some essential amino acids is insufficient probenecid reduces the transfer of benzylpenicillin into
to account for their output in milk protein (135). For ovine milk (266). Evidence for an organic anion transporter
example, in lactating goats, L-phenylalanine and L-histi- in mammary tissue is strengthened by the finding that an
dine uptake were found to be insufficient for the output of analog of p-aminohippuric acid (N4-acetylated p-aminohip-
these residues in milk. (8). On the basis that these amino puric acid), a model substrate of the kidney organic anion
acids cannot be synthesized within mammary cells, it has transport mechanism, is concentrated in bovine and caprine

include glutathione (␥-glutamyl-cysteinylglycine) and cer-

tain amino acids, is involved in amino acid transport. It
was envisaged that ␥-GT (in conjunction with other en-
zymes) utilized intracellular glutathione to effect the
transport of extracellular amino acids into the cell. How-
ever, the original model was revised on the finding that
the enzymatic site for glutathione was on the extracellular
face (80). The model predicts that following the hydroly-
sis of extracellular glutathione, the ␥-glutamyl moiety is
transferred to an amino acid to form a ␥-glutamyl-dipep-
tide that is subsequently taken up by cells. The initial
hypothesis that ␥-GT was involved in amino acid trans-
port was based on the correlation between high rates of
amino acid transport and ␥-GT activity. In this connec-
tion, lactating bovine and rat mammary tissue express
␥-GT activity with a specific activity second only to that
FIG. 6. Proposed scheme for the extracellular hydrolysis of anionic
dipeptides (using L-Glu-L-Ala as an example) and subsequent uptake of found in the kidney (19). In the rat, ␥-GT activity increases
the liberated anionic amino acid via the high-affinity, Na⫹-dependent with the onset of lactation and is regulated by prolactin. A
anionic amino acid carrier in lactating mammary cells. [From Shennan et role for ␥-GT in rat mammary tissue amino acid transport
al. (199). Copyright 1998, with permission from Elsevier Science.]
was inferred from the finding that the arteriovenous con-
centration gradients for those amino acids that are good
substrates of ␥-GT were reduced after the administration
milk (179). Active transport of 4-aminoantipyrine, N4-acety-
of inhibitors (serine borate or anthglutin) of ␥-GT (233,
lated sulfanilamide, and nitrofurantoin across the bovine,
235). However, interpretation of these results relies
caprine, and rat mammary gland, respectively, has also been
heavily on the assumption that the inhibitors are specific
described (10, 101, 102, 180, 227). There is the possibility that
for ␥-GT. The hypothesis that ␥-GT has a direct role in
an organic anion transport system, analogous to the one
amino acid transfer is weakened in light of the finding that
expressed in the kidney, may provide a route for small
the turnover of glutathione required to sustain amino acid
peptides across the basolateral membrane of mammary se-
transport is much higher than the predicted capacity of
cretory cells.
mammary tissue (236). An alternative explanation for the
The rat mammary gland is able to hydrolyze peptides
effect of ␥-GT on amino acid metabolism is that ␥-GT
extracellularly followed by uptake of the free amino acids
produces a signal that is capable of regulating amino acid
(199). Thus anionic peptides, presented to mammary tis-
uptake by rat mammary tissue. The main candidates are
sue explants or the perfused gland, are able to trans-
␥-glutamyl-amino acids or 5-oxoproline; the latter com-
stimulate D-aspartate efflux. It is envisaged that anionic
pound is formed intracellularly during the metabolism of
amino acids, produced as a consequence of peptidases
␥-glutamyl-amino acids (236, 238). Treating rats with bro-
that hydrolyze anionic dipeptides extracellularly, stimu-
mocriptine to reduce prolactin secretion inhibited ␥-GT
late D-aspartate efflux via the high-affinity anionic amino
⫺ and decreased the arteriovenous concentration difference
acid carrier, system XAG (199) (Fig 6). It appears that the
of a number of amino acids. However, amino acid extrac-
rat mammary gland has a large capacity to hydrolyze
tion by the rat mammary gland was restored to control
anionic peptides, since some dipeptides were almost as
levels by a product of ␥-GT, namely, ␥-glutamyl-glu-
effective as free L-glutamate in eliciting an increase in
tamine. Similarly, 5-oxoproline was shown to reverse the
D-aspartate efflux. It is possible that the uptake of free
bromocriptine-induced decrease in amino acid uptake by
amino acids following the extracellular hydrolysis of plas-
the rat mammary gland.
ma-derived peptides could make an important contribu-
Kansal and Kansal (99) have cast doubt on a func-
tion to the supply of certain amino acids for milk protein
tional relation between the activity of ␥-GT and amino
synthesis. The exact identity of the peptidases and the
acid uptake by the lactating mammary gland. They found
quantitative importance of blood peptides, as opposed to
that inhibition of ␥-GT had no effect on the uptake of
amino acids, remain to be established.
L-methionine and L-alanine by lactating mouse mammary
tissue explants.
F. Does ␥-Glutamyl Transpeptidase Play a Role Bovine mammary tissue ␥-GT is able to hydrolyze
in Mammary Amino Acid Transport? extracellular glutathione; the liberated amino acids were
subsequently transported into the intracellular compart-
Meister (128) proposed that ␥-glutamyl transpepti- ment (20). This observation coupled with the finding that
dase (␥-GT; EC 2.3.2.2.), an enzyme whose substrates there is an arteriovenous glutathione concentration dif-

ference across the bovine mammary gland suggested that local factors control that expression and activity after
glutathione, derived from red blood cells, may be an transcription, a major topic for future research must be
important source of cysteine for milk protein synthesis how all the intracellular events are integrated, whether by
(173). Similarly, it has been concluded that the goat mam- quantitatively coordinated gene expression in response to
mary gland also utilizes erythrocyte-derived glutathione hormonal or local signals or by cross-talk and feedback
as a source of amino acids (6). between the various pathways. In this respect, it is inter-
esting to note that the majority of work on intracellular
signaling within the mammary gland has been concerned
XIV. FATTY ACID UPTAKE
with mammary development, because of the obvious re-
lation to breast cancer, rather than with the control of
The amount of fat in milk, the majority of which is
secretion (250, 251).
triacylglycerol, ranges from 0.2% in rhinos to ⬎50% in
It is time for investigations of the coordinated regu-
pinnipeds (151). Lactating mammary tissue is able to
lation to extend to those changes in the composition of
synthesize fatty acids intracellularly from a supply of
the secretion that occur with physiological state, the for-
substrates extracted from plasma. In addition, fatty acids
mation of colostrum in late pregnancy, and the shift to
are extracted from lipids in the blood. The relative impor-
milk production at around the time of parturition, for
tance of intracellular synthesis compared with plasma-
example. There are also several major changes in milk
derived fatty acids depends on species, stage of lactation,
composition during lactation in marsupials, matching the
and diet (16, 115, 141). The plasma sources of fatty acids
requirements of the young at different stages in develop-
include triacylglycerols, cholesterol esters, and nonesteri-
ment (76, 84) that should receive increased attention.
fied fatty acids; quantitatively, triacylglycerols carried in
chylomicrons and very-low-density lipoproteins are the Address for reprint requests and other correspondence:
most important in fed animals. Nonesterified fatty acids D. B. Shennan, Hannah Research Institute, Ayr, Scotland KA6
are, however, important for mammary lipid uptake in the 5HL (E-mail: shennand@hri.sari.ac.uk and/or peakerm@hri.
fasting state (148). Nonesterified fatty acids are produced sari.ac.uk).
from triacylglycerols in mammary tissue by the enzyme
lipoprotein lipase which is situated in the capillary endo-
thelium (and possibly the basolateral surface of the se- REFERENCES
cretory cells). The mechanism of fatty acid uptake by 1. ABUMRAD NA, EL-MAGHRABI MR, AMRI E-Z, LOPEZ E, AND
mammary epithelial cells has not been identified. How- GRIMALDI PA. Cloning of a rat adipocyte membrane protein im-
ever, it has been suggested that fatty acid binding pro- plicated in binding or transport of long chain fatty acids that is
induced during preadipocyte differentiation. J Biol Chem 268:
teins, located in the plasma membrane, may play a role in 17665–17668, 1993.
fatty acid uptake (reviewed by Barber et al., Ref. 12). 2. AICKIN CC AND BRADING AF. Effect of Na⫹ and K⫹ on Cl⫺
Expression of a protein termed FAT (fatty acid translo- distribution in guinea-pig vas deferens smooth muscle: evidence for
Na⫹, K⫹, Cl⫺ co-transport. J Physiol (Lond) 421: 13–32, 1990.
cator) in fibroblasts induces the uptake/binding of long- 3. ALLEN JC. Sodium and potassium content and viability of mouse
chain fatty acids (91). FAT, which is predicted to have two mammary gland tissue and acini. J Dairy Sci 71: 633– 642, 1988.
membrane-spanning domains, may be either a fatty acid 4. AMATO PA AND LOIZZI RF. The effects of cytochalasin B on
glucose transport and lactose synthesis in lactating mammary
transporter or a regulator of fatty acid transport (1). gland slices. Eur J Cell Biol 20: 150 –155, 1979.
Interestingly, FAT has 85% homology with CD36, a protein 5. ATKINSON SA, SCHNURR CM, DONOVAN SM, AND LÖNNERDAL
expressed in the lactating mammary epithelium (77). B. The non-protein nitrogen components in human milk: biochem-
istry and potential functional role. In: Protein and Non-protein
Nitrogen in Human Milk, edited by S A. Atkinson and B. Lönner-
dal. Boca Raton, FL: CRC, 1989, p. 117–133.
XV. COORDINATION AND CONTROL 6. ATROSHI F, SANKARI S, POSO H, AND SANDHOLM M. Uptake of
blood amino acids and erythrocyte glutathione by the caprine
It is clear from this review of the transport mecha- mammary gland. J Anim Physiol Anim Nutr 55: 208 –213, 1986.
7. BACKWELL FRC, BEQUETTE BJ, WILSON D, CALDER AG, MET-
nisms operating across the basolateral membrane of the CALF JA, WRAY-COHEN D, MACRAE JC, BEEVER DE, AND LOB-
secretory cell that the degree of coordination between LEY GE. Utilization of dipeptides by the caprine mammary gland
synthetic, secretory, and transport processes to produce for milk protein synthesis. Am J Physiol Regulatory Integrative
Comp Physiol 267: R1–R6, 1994.
milk of virtually constant but complex composition must 8. BACKWELL FRC, BEQUETTE BJ, WILSON D, METCALF JA,
be very high. Even for protein synthesis, a key question is FRANKLIN MF, BEEVER DE, LOBLEY GE, AND MACRAE JC. Evi-
how the numerous transport processes for amino acids dence for the utilization of peptides for milk protein synthesis in
the lactating dairy goat in vivo. Am J Physiol Regulatory Integra-
act in concert to deliver the fixed-proportion mixture tive Comp Physiol 271: R955–R960, 1996.
required. 9. BANDERALI U AND ROY G. Anion channels for amino acids in
While efforts will continue to identify which trans- MDCK cells. Am J Physiol Cell Physiol 263: C1200 –C1207, 1992.
10. BANERJEE NC, MILLER GE, AND STOWE CM. Excretion of amino-
porters of those available within the genome are ex- pyrine and its metabolites into cows milk. Toxicol Appl Pharmacol
pressed by the mammary gland and what systemic and 10: 604 – 612, 1967.

11. BANNAI S AND KITAMURE E. Transport interaction of L-cystine J. Glucose transporter expression in rat mammary gland. Biochem
and L-glutamate in human diploid fibroblasts in culture. J Biol J 270: 277–279, 1990.
Chem 255: 2372–2376, 1980. 36. BUSTAMANTE JC, YUDILEVICH DL, AND BOYD CAR. A new form
12. BARBER MC, CLEGG RA, TRAVERS MT, AND VERNON RG. Lipid of asymmetry in epithelia: kinetics of apical and basal sulphate
metabolism in the lactating mammary gland. Biochim Biophys transport in human placenta. Q J Exp Physiol 73: 1013–1016, 1988.
Acta 1347: 101–126, 1997. 37. CABANTCHIK ZI AND GREGER R. Chemical probes for anion trans-
13. BARCELLOS-HOFF MH, AGGELO J, RAM TG, AND BISSELL MJ. porters of mammalian cell membranes. Am J Physiol Cell Physiol
Functional differentiation and alveolar morphogenesis of primary 262: C803–C827, 1992.
mammary cultures on reconstituted basement membrane. Devel- 38. CALVERT DT AND SHENNAN DB. Evidence for an interaction
opment 105: 223–235, 1989. between cationic and neutral amino acids at the blood-facing as-
14. BARKER GA AND ELLORY JC. The identification of neutral amino pect of the lactating rat mammary epithelium. J Dairy Res 63:
acid transport systems. Exp Physiol 75: 3–26, 1990. 25–33, 1996.
15. BARLET J-P, CHAMPREDON C, COXAM V, DAVICCO MJ, AND 39. CALVERT DT AND SHENNAN DB. Volume-activated taurine efflux
TRESSOL JC. Parathyroid hormone-related peptide might stimu- from the in situ perfused lactating rat mammary gland. Acta
late calcium secretion into the milk of goats. J Endocrinol 132: Physiol Scand 162: 97–105, 1998.
353–359, 1992. 40. CAMPS M, VILARO S, TESTAR X, PALACIN M, AND ZORZANO A.
16. BAUMAN DE AND DAVIS CL. Biosynthesis of milk fat. In: Lacta- High and polarized expression of GLUT1 glucose transporters in
tion, edited by B. L. Larson and V. R. Smith. New York: Academic, epithelial cells from mammary gland: acute downregulation of
1974, p. 31–75. GLUT1 carriers by weaning. Endocrinology 134: 924 –934, 1994.
17. BAUMRUCKER CR. Cationic amino acid transport by bovine mam- 41. CHAO CK, POMFRET EA, AND ZEISEL SH. Uptake of choline by rat
mary tissue. J Dairy Sci 67: 2500 –2506, 1984. mammary-gland epithelial cells. Biochem J 254: 33–38, 1988.
18. BAUMRUCKER CR. Amino acid transport systems in bovine mam- 42. CHEN L-H AND BISSELL MJ. A novel regulatory mechanism for
mary tissue. J Dairy Sci 68: 2436 –2451, 1985. whey acidic protein gene expression. Cell Regul 1: 45–54, 1989.
19. BAUMRUCKER CR AND POCIUS PA. ␥-Glutamyl transpeptidase in 43. CHIPPERFIELD AR. The (Na⫹K⫹Cl⫺) cotransport system. Clin Sci
lactating in lactating mammary secretory tissue of cow and rat. J (Lond) 71: 465– 476, 1986.
Dairy Sci 61: 309 –314, 1978. 44. CHRISTIE WW AND WOODING FBP. The site of triglyceride bio-
20. BAUMRUCKER CR, POCIUS PA, AND RISS TL. Glutathione utiliza- synthesis in milk. Experientia 31: 1445–1447, 1975.
tion by bovine mammary secretory tissue in vitro. Biochem J 198: 45. CLARK JH, SPIRES HR, AND DAVIS CL. Uptake and metabolism of
243–246, 1981. nitrogenous components by the lactating mammary gland. Feder-
21. BERNER W, KINNE R, AND MURER H. Phosphate transport into ation Proc 37: 1233–1238, 1978.
brush-border membrane vesicles isolated from rat small intestine. 46. CROSS BA. Comparative physiology of milk removal. In: Compar-
Biochem J 160: 467– 474, 1976. ative Aspects of Lactation, edited by M Peaker. London: Academic,
22. BERTRAN J, WERNER A, STRANGE G, MARKOVICH D, BIBER J, 1977, p. 193–210. (Symp Zool Soc Lond 41)
TESTAR X, ZORZANO A, PALACIN M, AND MURER H. Expression 47. DAI G, LEVY O, AND CARRASCO N. Cloning and characterization of
of a Na⫹-independent amino acid transport in Xenopus laevis the thyroid iodide transporter. Nature 379: 458 – 460, 1996.
oocytes by injection of rabbit kidney cortex mRNA. Biochem J 281: 48. DALY SEJ, KENT JC, OWENS RA, AND HARTMANN PE. Frequency
717–723, 1992. and degree of milk removal and the short-term control of human
23. BIBER J AND MURER H. Renal epithelial apical Na/Pi cotransport- milk synthesis. Exp Physiol 81: 861– 875, 1996.
ers. Cell Physiol Biochem 4: 185–197, 1994. 49. DAVIS AJ, FLEET IR, GOODE JA, HAMON MH, MAULE WALKER
24. BISBEE CA. Prolactin effects on ion transport across cultured FM, AND PEAKER M. Changes in mammary function at the onset of
mouse mammary epithelium. Am J Physiol Cell Physiol 240: C110 – lactation in the goat: correlation with hormonal changes. J Physiol
C115, 1981. (Lond) 288: 33– 44, 1979.
25. BISBEE CA, MACHEN TS, AND BERN HA. Mouse mammary epi- 50. DAVIS JPL. The effect of (Na-K-Cl) cotransport and Cl-HCO3 ex-
thelial cells on floating collagen gels: transepithelial ion transport change blockade on the membrane potential and intracellular chlo-
and effects of prolactin. Proc Natl Acad Sci USA 76: 536 –540, 1979. ride levels of rat arterial smooth muscle, in vitro. Exp Physiol 77:
26. BISBEE CE. Transepithelial electrophysiology of cultured mouse 857– 862, 1992.
mammary epithelium: sensitivity to prolactins. Am J Physiol En- 51. DAVIS-AMARAL E, MUSCH MW, AND GOLDSTEIN L. Chloride and
docrinol Metab 241: E410 –E413, 1981. taurine effluxes occur by different pathways in skate erythrocytes.
27. BLATCHFORD DR AND PEAKER M. Effect of ionic composition of Am J Physiol Regulatory Integrative Comp Physiol 271: R1544 –
milk on transepithelial potential in the goat mammary gland. R1549, 1996.
J Physiol (Lond) 402: 533–541, 1988. 52. DEVES R, CHAVEZ P, AND BOYD CAR. Identification of a new
28. BOESE SH, WEHNER F, AND KINNE RH. Taurine permeation transporter system (y⫹L) in human erythrocytes that recognises
through swelling-activated anion conductance in rat IMCD cells in lysine and leucine with high affinity. J Physiol (Lond) 454: 491–501,
primary culture. Am J Physiol Renal Fluid Electrolyte Physiol 271: 1992.
F498 –F507, 1996. 53. DJURDJEVIC DJ AND LENGEMANN FW. Effect of perchlorate on
29. BRETON S, MARSOLAIS M, AND LAPRADE R. Hypotonicity in- the milk plasma radioiodine ratio of intact and thyroidectomised
creases basolateral taurine permeability in rabbit proximal convo- goats. J Dairy Sci 53: 808 – 812, 1970.
luted tubule. Am J Physiol Renal Fluid Electrolyte Physiol 268: 54. DORING F, WALTER J, WILL J, FOCKING M, BOLL M, AMASHEH
F595–F603, 1995. S, CLAUSS W, AND DANIEL H. Delta-aminolevulinic acid transport
30. BROMMAGE R. Measurement of calcium and phosphorus fluxes by intestinal and renal peptide transporters and its physiological
during lactation in the rat. J Nutr 119: 428 – 438, 1988. and clinical implications. J Clin Invest 101: 2761–2767, 1998.
31. BROWN-GRANT K. The iodide concentration mechanism of the 55. DOWBEN RM, BRUNNER JR, AND PHILPOTT DE. Studies on milk
mammary gland. J Physiol (Lond) 135: 644 – 654, 1957. fat globule membranes. Biochim Biophys Acta 135: 1–10, 1967.
32. BROWN-GRANT K. Extrathyroidal iodide concentrating mecha- 56. DUNCAN JS AND BURGOYNE RD. Characterization of the effects
nisms. Physiol Rev 41: 189 –213, 1961. of Ca2⫹ depletion on the synthesis, phosphorylation and secretion
33. BRUNETTE MG AND ALLARD S. Phosphate uptake by syncytial of caseins in lactating mammary epithelial cells. Biochem J 317:
brush-border membranes of the human placenta. Pediatr Res 19: 487– 493, 1996.
1179 –1182, 1985. 57. ENDERS RH, JUDD JM, DONOHUE TM, AND SMITH CH. Placental
34. BURNOL AF, FERRE P, LETURQUE A, AND GIRARD J. Effect of amino acid uptake. III. Transport systems for neutral amino acids.
insulin on in vivo glucose utilization in individual tissues of anaes- Am J Physiol 230: 706 –710, 1976.
thetized lactating rats. Am J Physiol Endocrinol Metab 252: E183– 58. ENOMOTO K, FURUYA K, MAENO T, EDWARDS C, AND OKA T.
E188, 1987. Mechanically induced electrical responses in murine mammary
35. BURNOL AF, LETIRQUE A, LOIZEAU M, POSTIC C, AND GIRARD epithelial cells in primary culture. FEBS Lett 223: 82– 86, 1987.

59. ENOMOTO K, FURUYA K, MAENO T, EDWARDS C, AND OKA T. lyte transport in HeLa cells. Am J Physiol Cell Physiol 271: C579 –
Oscillating activity of a calcium-activated K⫹ channel in normal C588, 1996.
and cancerous mammary cells in culture. J Membr Biol 119: 133– 82. HANIGAN MD, CALVERT CC, DEPETERS EJ, REIS BL, AND BALD-
139, 1991. WIN RL. Kinetics of amino acid extraction by lactating mammary
60. ENOMOTO K, FURUYA K, YAMAGISHI S, AND MAENO T. Mechan- glands in control and sometribove-treated Holstein cows. J Dairy
ically induced electrical and intracellular calcium responses in Sci 75: 161–173, 1992.
normal and cancerous mammary cells. Cell Calcium 13: 501–511, 83. HANWELL A AND PEAKER M. Physiological effects of lactation on
1992. the mother. In: Comparative Aspects of Lactation, edited by M.
61. FALCONER IR, LANGLEY JV, AND VACEK AT. Effect of prolactin Peaker. London: Academic, 1977, p. 297–312.
on 86Rb uptake, potassium content and [G-3H]ouabain binding of 84. HINDS LA. Hormonal control of lactation. In: The Developing
lactating rabbit mammary tissue. J Physiol (Lond) 334: 1–17, 1983. Marsupial, edited by C. H. Tyndale-Biscoe and P. A. Janssens.
62. FALCONER IR AND ROWE JM. Possible mechanism for action of Heidelberg, Germany: Springer-Verlag, 1988, p. 55– 67.
prolactin on mammary cell sodium transport. Nature 256: 327–328, 85. HOFFMAN E AND SIMONSEN LO. Membrane mechanisms in vol-
1975. ume and pH regulation in vertebrate cells. Physiol Rev 69: 315–382,
63. FALCONER IR AND ROWE JM. Effect of prolactin on sodium and 1989.
potassium concentrations in mammary alveolar tissue. Endocrinol- 86. HOLT C AND JENNESS R. Interrelationships of constituents and
ogy 101: 181–186, 1977. partition of salts in milk samples from eight species. Comp Bio-
64. FAULKNER A, BLATCHFORD DR, AND POLLOCK HT. The trans- chem Physiol A Physiol 77: 275–282, 1984.
port of hexoses across the apical membrane of the mammary gland 87. HUDSON ER, MA L, WILDE CJ, FLINT DJ, AND BALDWIN SA.
of the goat. Biochem Soc Trans 13: 689 – 690, 1985. Regulation of GLUT1 expression in the mammary gland. Biochem
65. FAULKNER A, CHAIYABUTR N, PEAKER M, CARRICK DT, AND Soc Trans 25: 92, 1997.
KUHN NJ. The metabolic significance of milk glucose. J Dairy Res 88. HUNZIKER W, AND KRAEHENBUHL J-P. Epithelial transcytosis of
48: 51–56, 1981. immunoglobulins. J Mammary Gland Biol Neoplasia 3: 287–302,
66. FAWCETT HAC, BALDWIN SA, AND FLINT DJ. Hormonal regula- 1998.
tion of the glucose transporter GLUT 1 in the lactating rat mam- 89. HURLEY WL, BLATCHFORD DR, HENDRY KAK, AND WILDE CJ.
mary gland (Abstract). Biochem Soc Trans 20: 17S, 1991. Extracellular matrix and mouse mammary cell function: compari-
67. FEI Y-J, KANAI Y, NUSSBERGER S, GANAPATHY V, LEIBACH FH, son of substrata in culture. In Vitro 30: 529 –538, 1994.
ROMERO MF, SINGH SK, BORON WF, AND HEDIGER MA. Expres- 90. HUXTABLE RJ. Physiological actions of taurine. Physiol Rev 72:
sion cloning of a mammalian proton-coupled oligopeptide trans- 101–163, 1992.
porter. Nature 368: 563–566, 1994. 91. IBRAHIMI A, SFEIR Z, MAGHARAIE H, AMRI EZ, GRIMALDI P,
AND ABUMRAD NA. Expression of the CD36 homolog (FAT) in
68. FLEET IR, GOODE JA, HAMON MH, LAURIE MS, LINZELL JL, AND
PEAKER M. Secretory activity of goat mammary glands during fibroblast cells: effects on fatty acid transport. Proc Natl Acad Sci
USA 93: 2646 –2651, 1996.
pregnancy and the onset of lactation. J Physiol (Lond) 252: 763–
92. JACKSON PS AND STRANGE K. Volume-sensitive anion channels
773, 1975.
mediate swelling-activated inositol and taurine efflux. Am J
69. FLEET IR AND PEAKER M. Mammary function and its control at the
Physiol Cell Physiol 265: C1489 –C1500, 1993.
cessation of lactation in the goat. J Physiol (Lond) 279: 491–507,
93. JANSEN GR, SCHILBY MB, MASOR M, SAMPSON DA, AND LON-
1978.
GENECKER JB. Free amino acid levels during lactation in rats:
70. FLEEZER M AND HEISLER S. P2-purinergic receptors in human
effects of protein quantity and quality. J Nutr 116: 376 –387, 1986.
breast tumor cells: coupling of intracellular calcium signaling to
94. JENNESS R. The composition of milk. In: Lactation, edited by B L.
anion secretion. J Pharmacol Exp Ther 265: 1499 –1510, 1993.
Larson and V. R. Smith. New York: Academic, 1974, p. 3–107.
71. FLINT DJ AND GARDNER M. Evidence that growth hormone stim-
95. JESSEN H. Taurine and ␤-alanine transport in an established hu-
ulates milk synthesis by direct action on the mammary gland and man kidney cell line derived from the proximal tubule. Biochim
that prolactin exerts effects on milk secretion by maintenance of Biophys Acta 1194: 44 –52, 1994.
mammary DNA content and tight junction status. Endocrinology 96. JOHNSON MP AND WOODING FBP. Adenosine triphosphatase dis-
135: 1119 –1124, 1994. tribution in mammary tissue. Histochem J 10: 171–183, 1978.
72. FURUYA K. Single calcium-activated potassium channel in cul- 97. KANAI Y AND HEDIGER MA. Primary structure and functional
tured mammary epithelial cells. Pflügers Arch 414: 118 –124, 1989. characterization of a high affinity glutamate transporter. Nature
73. FURUYA K, ENOMOTO K, MAENO T, AND YAMAGISHI S. Mechan- 360: 467– 471, 1992.
ically induced calcium signal in mammary epithelial cells. Jpn 98. KANAI Y, SMITH GP, AND HEDIGER MA. A new family of neuro-
J Physiol 43 Suppl: S105–S108, 1993. transmitter transporters: the high affinity glutamate transporters.
74. GOULD GW AND HOLMAN GD. The glucose transporter family: FASEB J 8: 1450 –1459, 1993.
structure function and tissue-specific expression. Biochem J 295: 99. KANSAL R AND KANSAL K. Lack of functional correlation between
329 –341, 1993. gamma-glutamyl transpeptidase and amino acid transport in the
75. GOW IF, BURNS C, AND SHENNAN DB. Volume-activated calcium lactating mouse mammary gland. Indian J Exp Biol 34: 267–269,
transport in lactating rat mammary tissue (Abstract). Biochem Soc 1996.
Trans 26: 389S, 1998. 100. KANSAL R AND KANSAL VR. Discrimination of transport systems
76. GREEN B AND MERCHANT JC. The composition of marsupial milk. for L-tyrosine in mouse mammary gland: characterisation of system
In: The Developing Marsupial, edited by C H. Tyndale-Biscoe and T. Indian J Exp Biol 34: 750 –757, 1996.
P. A. Janssens. Heidelberg, Germany: Springer-Verlag, 1988, p. 41– 101. KARI FW, WEAVER R, AND NEVILLE MC. Active transport of
54. nitrofurantoin across a mouse mammary epithelial monolayer.
77. GREENWALT DE, LIPSKY RH, OCKENHOUSE CF, IKEDA H, TAN- J Pharmacol Exp Ther 280: 664 – 668, 1997.
DON NN, AND JAMIESON GA. Membrane glycoprotein CD36: a 102. KARL FW, WEAVER R, AND NEVILLE MC. Active transport of
review of its role in adherence, signal transduction, and transfusion nitrofurantoin across the mammary epithelium in vivo. J Pharma-
medicine. Blood 80: 1105–1115, 1992. col Exp Ther 280: 664 – 668, 1997.
78. GROSVENOR CE, PICCIANO MF, AND BAUMRUCKER CR. Hor- 103. KEMPSON SA. Novel specific inhibitors of epithelial phosphate
mones and growth factors in milk. Endocr Rev 14: 710 –728, 1992. transport. News Physiol Sci 3: 154 –157, 1988.
79. HAAS M. The Na-K-Cl cotransporters. Am J Physiol Cell Physiol 104. KILBERG MS, HANDLOGTEN ME, AND CHRISTENSEN HN. Char-
267: C869 –C885, 1994. acteristics of an amino acid transport system in rat liver for glu-
80. HAHN RA, WENDEL A, AND FLOHE L. The fate of extracellular tamine, asparagine, histidine and closely related analogs. J Biol
glutathione in the rat. Biochim Biophys Acta 539: 324 –337, 1978. Chem 255: 4011– 4019, 1980.
81. HALL JA, KIRK J, POTTS JR, RAE C, AND KIRK K. Anion channel 105. KINURA T. Electron microscopic study of the mechanism of se-
blockers inhibit swelling-activated anion, cation and non-electro- cretion of milk. J Jpn Obstet Gynecol Soc 21: 301–308, 1969.

106. KIRK K. Swelling-activated organic osmolyte channels. J Membr parative Aspects of Lactation, edited by M. Peaker. London: Aca-
Biol 158: 1–16, 1997. demic, 1977, p. 57–75.
107. KIRK K, ELLORY JC, AND YOUNG JD. Transport of organic sub- 130. MEPHAM TB AND LINZELL JL. Urea formation of the lactating goat
strates via a volume-activated channel. J Biol Chem 267: 23475– mammary gland. Nature 214: 507–508, 1967.
23468, 1992. 131. MEPHAM TB, OVERTHROW JI, AND SHORT AH. Epithelial cell
108. KIRK K AND KIRK J. Volume-regulatory taurine release from a entry and exit competition amongst amino acids in the isolated
human lung cancer cell line. Evidence for amino acid transport via perfused lactating mammary gland of the guinea pig. In: Carrier
a volume-activated chloride channel. FEBS Lett 336: 153–158, 1993. Mediated Transport of Solutes From Blood to Tissue, edited by
109. KLOCKNER U, STORCK T, CONRADT M, AND STOFFEL W. Func- D. L. Yudilevich and G. E. Mann. London: Longman, 1985, p. 369 –
tional properties and substrate specificity of the cloned L-gluta- 372.
mate/L-aspartate transporter GLAST-1 from rat brain expressed in 132. MEREDITH D AND BOYD CAR. Oligopeptide transport by epithelial
Xenopus oocytes. J Neurosci 14: 5759 –5765, 1994. cells. J Membr Biol 145: 1–12, 1995.
110. KRONFELD DS. Major metabolic determinants of milk volume, 133. MESSER M AND ELLIOTT C. Changes in ␣-lactalbumin, total lac-
mammary efficiency, and spontaneous ketosis in dairy cows. J tose, UDP-galactose hydrolase and other factors in Tammar wal-
Dairy Sci 65: 2204 –2212, 1982. laby (Macropus eugenii) milk during lactation. Aust J Biol Sci 40:
111. KULPA HU. Whole cell chloride currents in cultivated mammary 37– 46, 1987.
epithelial cells (Abstract). Pflugers Arch 433: 590, 1997. 134. MESSER M AND GREEN B. Milk carbohydrate of marsupials. II.
112. LACASSE P, FARR VC, DAVIS SR, AND PROSSER CG. Local secre- Quantitative and qualitative changes in milk carbohydrates during
tion of nitric oxide and the control of mammary blood flow. J lactation in the tammar wallaby (Macropus eugenii). Aust J Biol
Dairy Sci 79: 1369 –1374, 1996. Sci 32: 519 –531, 1979.
113. LAMBERT IH AND HOFFMANN EK. Cell-swelling activates sepa- 135. METCALF JA, BEEVER DE, SUTTON JD, WRAY-CAHEN D,
rate taurine and Cl⫺ channels in Ehrlich mouse ascites tumor cells. EVANS RT, HUMPHRIES DJ, BACKWELL FRC, BEQUETTE BJ,
J Membr Biol 142: 289 –298, 1994. AND MACRAE JC. The effect of supplementary protein on in vivo
114. LENGEMANN FW. Reduction of iodide transfer to milk of cows metabolism of the mammary gland in lactating dairy cows. J Dairy
after perchlorate ingestion. J Dairy Sci 56: 753–756, 1973. Sci 77: 1816 –1827, 1994.
115. LINZELL JL. Mammary blood flow and methods of identifying and 136. METCALF JA, SUTTON JD, COCKBURN JE, NAPPER DJ, AND
measuring precursors of milk. In: Lactation, edited by B. L. Larson BEEVER DE. The influence of insulin and amino acid supply on
and V. R. Smith. New York: Academic, 1974, p. 143–225. amino acid uptake by the lactating bovine mammary gland. J Dairy
116. LINZELL JL, MEPHAM TB, AND PEAKER M. The secretion of Sci 74: 3412–3420, 1991.
citrate into milk. J Physiol (Lond) 260: 739 –750, 1976. 137. MEZZETTI G, MONTI MG, CASOLO LP, PICCININI G, AND
117. LINZELL JL AND PEAKER M. Intracellular concentrations of so- MORUZZI MS. 1,25-Dihydroxycholecalciferol-dependent calcium
dium, potassium and chloride in the lactating mammary gland and uptake by mouse mammary gland in culture. Endocrinology 122:
389 –394, 1988.
their relation to the secretory mechanism. J Physiol (Lond) 216:
138. MILLAR ID, CALVERT DT, LOMAX MA, AND SHENNAN DB. The
683–700, 1971.
mechanism of L-glutamate transport by lactating rat mammary
118. LINZELL JL AND PEAKER M. Mechanism of milk secretion. Physiol
tissue. Biochim Biophys Acta 1282: 200 –206, 1996.
Rev 51: 564 –597, 1971.
139. MILLAR ID, CALVERT DT, LOMAX MA, AND SHENNAN DB. Sub-
119. LINZELL JL AND PEAKER M. Changes in colostrum composition
strate specificity of the mammary tissue anionic amino acid carrier
and in the permeability of the mammary epithelium at about the
operating in the cotransport and exchange modes. Biochim Bio-
time of parturition in the goat. J Physiol (Lond) 243: 129 –151, 1974.
phys Acta 1326: 92–102, 1997.
120. LINZELL JL AND PEAKER M. The distribution and movements of
140. NALECZ KA, KORZON D, WAWRZENCZYCK A, AND NALECZ MJ.
carbon dioxide, carbonic acid and bicarbonate between blood and
Transport of carnitine in neuroblastoma NB-2A cells. Arch Bio-
milk in the goat. J Physiol (Lond) 244: 771–782, 1975. chem Biophys 322: 214 –220, 1995.
121. LINZELL JL, PEAKER M, AND TAYLOR JC. The effects of prolactin 141. NEVILLE MC, ALLEN JC, AND WATTERS C. The mechanisms of
and oxytocin on milk secretion and on the permeability of the milk secretion. In: Lactation, edited by M. C. Neville and M. R.
mammary epithelium in the rabbit. J Physiol (Lond) 253: 547–563, Neifert. New York: Plenum, 1983, p. 49 –102.
1975. 142. NEVILLE MC, HAY WW, AND FENNESSEY P. Physiological impor-
122. LIU QR, LOPEZ-CORCERA B, NELSON H, MANDIYAN S, AND tance of the concentration of human-milk glucose. Protoplasma
NELSON H. Cloning and expression of a cDNA encoding the trans- 159: 118 –128, 1990.
porter of taurine and ␤-alanine in mouse brain. Proc Natl Acad Sci 143. NEVILLE MC, JACOBS D, AND EVANS T. Calcium transport in the
USA 89: 12145–12149, 1992. mammary gland: mechanism for maintaining low intracellular cal-
123. LIU W, LIANG S, RAMAMOORTHY S, FEI Y-J, GANAPATHY ME, cium in mammary tissue from lactating mice (Abstract). Federa-
HEDIGER MA, GANAPATHY V, AND LEIBACH FH. Molecular clon- tion Proc 39: 1966, 1980.
ing of PEPT2, a new member of the H⫹/peptide cotransporter 144. NEVILLE MC, KELLER RP, CASEY CE, AND ALLEN JC. Calcium
family, from human kidney. Biochim Biophys Acta 1235: 461– 466, partitioning in human and bovine milk. J Dairy Sci 77: 1964 –1975,
1995. 1994.
124. MADON RJ, MARTIN S, DAVIES A, FAWCETT HAC, FLINT DJ, AND 145. NEVILLE MC, LOBITZ CJ, RIPOLL EA, AND TINNEY C. The sites for
BALDWIN SA. Identification and characterization of glucose trans- ␣-aminoisobutyric acid uptake in normal mammary gland and as-
port proteins in plasma membrane- and Golgi vesicle-enriched cites tumor cells. J Biol Chem 255: 7311–7316, 1980.
fractions prepared from lactating rat mammary gland. Biochem J 146. NEVILLE MC AND PEAKER M. The secretion of calcium and phos-
272: 99 –105, 1990. phorus into milk. J Physiol (Lond) 290: 59 – 67, 1979.
125. MAGAGNIN S, WERNER A, MARKOVISH D, SORRIBAS V, 147. NEVILLE MC AND PEAKER M. Calcium fluxes in mouse mammary
STRANGE G, BIBER J, AND MURER H. Expression cloning of a tissue in vitro: intracellular and extracellular calcium pools.
human and rat renal-cortex Na⫹/Pi cotransporter. Proc Natl Acad J Physiol (Lond) 232: 497–517, 1982.
Sci USA 90: 5979 –5983, 1993. 148. NEVILLE MC AND PICCIANO MF. Regulation of milk lipid synthesis
126. MATHER IH AND KEENAN TW. Origin and secretion of milk lipids. and composition. Annu Rev Nutr 17: 159 –183, 1997.
JAMA 3: 259 –273, 1998. 149. NEVILLE MC, SELKER F, SEMPLE K, AND WATTERS C. ATP-
127. MAULE WALKER FM AND PEAKER M. Local production of pros- dependent calcium transport by a Golgi-enriched membrane frac-
taglandins in relation to mammary function at the onset of lactation tion from mouse mammary gland. J Membr Biol 61: 97–105, 1981.
in the goat. J Physiol (Lond) 309: 65–79, 1980. 150. NGUYEN D-AD AND NEVILLE MC. Tight junction regulation in the
128. MEISTER A. On the enzymology of amino acid transport. Science mammary gland. J Mammary Gland Biol Neoplasia 3: 227–246,
180: 33–39, 1973. 1998.
129. MEPHAM TB. Synthesis and secretion of milk proteins. In: Com- 151. OFTEDAL OT. Milk composition, milk yield and energy output at

peak lactation: a comparative review. In: Physiological Strategies LEIBACH FH, AND GANAPATHY V. Sodium-dependent carnitine
in Lactation, edited by M. Peaker, R. G. Vernon, and C. H. Knight. transport in human placental choriocarcinoma cells. Biochim Bio-
London: Academic, 1984, p. 33– 85. phys Acta 1284: 109 –117, 1996.
152. OFTEDAL OT, BONESS DJ, AND TEDMAN RA. The behavior, phys- 175. PROSSER CG. Mechanism of the decrease in hexose transport by
iology and anatomy of lactation in the Pinnipedia. In: Current mouse mammary epithelial cells caused by fasting. Biochem J 249:
Mammalogy, edited by H. H. Genoways. New York: Plenum, 1987, 149 –154, 1988.
p. 175–245. 176. PROSSER CG, DAVIS SR, FARR VC, AND LACASSE P. Regulation of
153. O’GRADY SM, PALFREY HC, AND FIELD M. Characteristics and blood flow in the mammary microvasculature. J Dairy Sci 79:
functions of Na-K-Cl co-transport in epithelial tissues. Am J 1184 –1197, 1996.
Physiol Cell Physiol 253: C177–C192, 1987. 177. PROSSER CG AND TOPPER YJ. Changes in the rate of carrier-
154. OLLIVIER-BOUSQUET M. Transferrin and prolactin transcytosis in mediated glucose transport by mouse mammary epithelial cells
the lactating mammary epithelial cell. J Mammary Gland Biol during ontogeny: hormone dependence delineated in vitro. Endo-
Neoplasia 3: 303–313, 1998. crinology 119: 91–96, 1986.
155. O’NEILL B, MAGNOLATO D, AND SEMENZA G. The electrogenic 178. RAMAMOORTHY S, LEIBACH FH, MAHESH VB, HAN H, YANG-
and Na-dependent iodide transport system in plasma membrane FENG T, BLAKELY RO, AND GANAPATHY V. Functional charac-
vesicles from thyroid glands. Biochim Biophys Acta 896: 263–274, terization and chromosome location of a cloned taurine transporter
1987. from human placenta. Biochem J 300: 893–900, 1994.
156. PAGE T AND KUHN NJ. Arteriovenous glucose differences across 179. RASMUSSEN F. Active mammary excretion of N4-acetylated p-
the mammary gland of fed, starved, and re-fed lactating rat. Bio- aminohippuric acid. Acta Vet Scand 10: 193–194, 1969.
chem J 239: 269 –274, 1986. 180. RASMUSSEN F. Active mammary excretion of N4-acetylated sul-
157. PASSANTES-MORALES H, MORAN J, AND SCHOUSBOE A. Vol- phanilamide. Acta Vet Scand 10: 402– 403, 1969.
ume-sensitive release of taurine from cultured astrocytes: proper- 181. RASSIN DK, STURMAN JA, AND GAULL GE. Taurine and other free
ties and mechanism. Glia 3: 427– 432, 1990. amino acids in milk of man and other mammals. Early Hum Dev 2:
158. PEAKER M. The aqueous phase of milk: ion and water transport. 1–13, 1978.
In: Comparative Aspects of Lactation, edited by M. Peaker. Lon- 182. REBOUCHE CJ AND MACK DL. Sodium gradient-stimulated trans-
don: Academic, 1977, p. 113–134. port of L-carnitine into renal brush-border membrane-vesicles—
159. PEAKER M. Mechanism of milk secretion: milk composition in kinetics, specificity and regulation by dietary carnitine. Arch Bio-
relation to potential difference across the mammary epithelium. chem Biophys 235: 393– 402, 1984.
J Physiol (Lond) 270: 489 –505, 1977. 183. RILLEMA JA, GOLDEN K, AND JENKINS MA. Effect of prolactin on
160. PEAKER M. Ion and water transport in the mammary gland. In: ␣-aminoisobutyric acid uptake in mouse mammary gland explants.
Lactation, edited by B. L. Larson. New York: Academic, 1978, p. Am J Physiol Endocrinol Metab 262: E402–E405, 1992.
437– 462. 184. RILLEMA JA AND ROWADY DL. Characteristics of the prolactin
161. PEAKER M. The effects of raised intramammary pressure on mam-
stimulation of iodide uptake into mouse mammary gland explants.
mary function in the goat in relation to the cessation of lactation.
Proc Soc Exp Biol Med 215: 366 –369, 1997.
J Physiol (Lond) 301: 415– 428, 1980.
185. RILLEMA JA AND YU TX. Prolactin stimulation of iodide uptake
162. PEAKER M. This symposium–with hindsight. In: Physiological
into mouse mammary gland explants. Am J Physiol Endocrinol
Strategies in Lactation, edited by M. Peaker, R. G. Vernon, and
Metab 271: E879 –E882, 1996.
C. H. Knight. London: Academic, 1984, p. xi-xiv.
186. ROBLES-VALDES C, MACGARRY D, AND FOSTER DW. Maternal-
163. PEAKER M. Autocrine control of milk secretion: development of
fetal carnitine relationships and neonatal ketosis in the rat. J Biol
the concept. In: Intercellular Signalling in the Mammary Gland.
Chem 251: 6007– 6012, 1976.
Proceedings of the Hannah Symposium, Ayr, 13–15 April 1994,
187. ROH SG, BAIK MG, AND CHOI YJ. The effect of lactogenic hor-
edited by C. J. Wilde, M. Peaker, and C. H. Knight. New York:
Plenum, 1995, p. 193–202. mones on protein synthesis and amino acid uptake in rat mammary
164. PEAKER M AND LINZELL JL. The effects of oestrus and exogenous acinar cell culture at various physiological stages. Int J Biochem
oestrogens on milk secretion in the goat. J Endocrinol 61: 231–240, 26: 479 – 485, 1994.
1974. 188. ROMAN RM, YANG Y, AND FITZ JG. Regulation of cell volume in a
165. PEAKER M AND NEVILLE MC. Hormones in milk: chemical signals human biliary cell line: activation of K⫹ and Cl⫺ currents. Am J
to the offspring. J Endocrinol 131: 1–3, 1991. Physiol Gastrointest Liver Physiol 271: G239 –G248, 1996.
166. PEAKER M AND TAYLOR E. Bumetanide binding to milk fat globule 189. ROSENBERG R, YOUNG JD, AND ELLORY JC. L-Tryptophan trans-
membrane (Abstract). J Physiol (Lond) 386: 75P, 1987. port in human red blood cells. Biochim Biophys Acta 598: 375–384,
167. PEAKER M AND TAYLOR JC. Milk secretion in the rabbit: changes 1980.
during lactation and the mechanism of ion transport. J Physiol 190. RUSSELL JM. Cation-coupled chloride influx in squid axon. Role of
(Lond) 253: 527–545, 1975. potassium and stoichiometry of the transport process. J Gen
168. PEAKER M AND WILDE CJ. Feedback control of milk secretion Physiol 81: 909 –925, 1983.
from milk. J Mammary Gland Biol Neoplasia 1: 307–315, 1996. 191. SANCHEZ-OLEA R, FULLER C, BENOS D, AND PASANTES-MO-
169. PEAKER M, WILDE CJ, AND KNIGHT CH. Local control of the RALES H. Volume-associated osmolyte fluxes in cell lines with or
mammary gland. In: Mammary Development and Cancer, edited without the anion exchanger. Am J Physiol Cell Physiol 269:
by P. S. Rudland, D. G. Fernig, S. Leinster and G. G. Lunt. London: C1280 –C1286, 1995.
Portland, 1997, p. 71–79. 192. SARWAR G, BOTTING HG, DAVIS TA, DARLING P, AND PEN-
170. PETERS BJ AND RILLEMA JA. Effect of prolactin on 2-deoxyglu- CHARZ PB. Free amino acids in milks of human subjects, other
cose uptake in mouse mammary gland explants. Am J Physiol primates and non-primates. Br J Nutr 79: 129 –131, 1998.
Endocrinol Metab 262: E627–E630, 1992. 193. SCHADEWINKEL-SCHERKL AM, RASMUSSEN F, MERCK C-C,
171. PINES G, DANBOLT NC, BJORAS M, ZHANG Y, BENDAHAN A, NIELSEN P, AND FREY H-H. Active transport of benzylpenicillin
EIDE L, KOEPSELL H, STORM-MATHISEN J, SEEBERG E, AND across the blood-milk barrier. Pharmacol Toxicol 73: 14 –19, 1993.
KANNER BL. Cloning and expression of a rat brain glutamate 194. SHENNAN DB. A study of sulphate transport by lactating rat
transporter. Nature 360: 464 – 467, 1992. mammary tissue slices: evidence for anion exchange. Comp Bio-
172. PLANTZ PE, PATTON S, AND KEENAN TW. Further evidence of chem Physiol A Physiol 92: 145–150, 1989.
plasma membrane material in skim milk. J Dairy Sci 56: 978 –983, 195. SHENNAN DB. Is the milk-fat-globule membrane a model for mam-
1973. mary secretory cell apical membrane? Exp Physiol 77: 653– 656,
173. POCIUS PA, CLARK JH, AND BAUMRUCKER CR. Glutathione in 1992.
bovine blood: possible source of amino acids for milk synthesis. J 196. SHENNAN DB. K⫹ and Cl- transport by mammary secretory cell
Dairy Sci 64: 1551–1554, 1981. apical membrane vesicles isolated from milk. J Dairy Res 59:
174. PRASAD PD, HUANG W, RAMAMOORTHY S, CARTER AL, 339 –348, 1992.

197. SHENNAN DB. Identification of a high affinity taurine transporter Am J Physiol Regulatory Integrative Comp Physiol 273: R379 –
which is not dependent on chloride. Biosci Rep 15: 231–239, 1995. R386, 1997.
198. SHENNAN DB AND BEECHEY RB. Mechanisms involved in the 218. STIPANUK MH, KUO S-M, AND HIRSCHBERGER LL. Changes in
uptake of D-glucose into the milk producing cells of rat mammary maternal taurine levels in response to pregnancy and lactation. Life
tissue. Biochem Biophys Res Commun 211: 986 –990, 1995. Sci 35: 1149 –1155, 1984.
199. SHENNAN DB, CALVERT DT, BACKWELL FRC, AND BOYD CAR. 219. STORCK T, SCHULTE S, HOFMANN K, AND STOFFEL W. Struc-
Peptide aminonitrogen transport by the lactating rat mammary ture, expression and functional analysis of a Na⫹-dependent gluta-
gland. Biochim Biophys Acta 1373: 252–260, 1998. mate aspartate transporter from rat brain. Proc Natl Acad Sci USA
200. SHENNAN DB, CLIFF MJ, AND HAWKINS P. Volume-sensitive tau- 89: 10959 –10965, 1992.
rine efflux is not obliged to utilize volume-activated anion channels. 220. STURMAN JA, RASSIN DK, AND GAULL GE. Taurine in developing
Biosci Rep 16: 459 – 465, 1996. rat brain: transfer of [35S]taurine to pups via the milk. Pediatr Res
201. SHENNAN DB, GRANT A, RAMSAY RR, BURNS C, AND ZAMMIT 11: 28 –33, 1977.
VA. Characteristics of L-carnitine transport by lactating rat mam- 221. SUDLOW AW AND BURGOYNE RD. A hypo-osmotically induced
mary tissue. Biochim Biophys Acta 1393: 49 –56, 1998. increase in intracellular Ca2⫹ in lactating mouse mammary epithe-
202. SHENNAN DB AND MADON RJ. An effect of prolactin on K uptake lial cells involving Ca2⫹ influx. Pflügers Arch 433: 609 – 616, 1997.
by lactating rat mammary tissue. Horm Metab Res 23: 293–294, 222. TAKATA K, FUJIKURA K, SUZUKI M, SUZUKI T, AND HIRANO H.
1991. GLUT1 glucose transporter in the lactating mammary gland in the
203. SHENNAN DB AND MCNEILLIE SA. Efflux of chloride from lactat- rat. Acta Histochem Cytochem 30: 623– 628, 1997.
ing rat mammary tissue. Comp Biochem Physiol A Physiol 95: 223. TEDSTONE AE, ILIC V, AND WILLIAMSON DH. Reciprocal changes
367–371, 1990. in amino acid metabolism in mammary gland and liver of the
204. SHENNAN DB AND MCNEILLIE SA. Characteristics of ␣-amino- lactating rat on starvation and refeeding as indicated by the tissue
isobutyric acid transport by lactating rat mammary gland. J Dairy accumulation of ␣-amino[1-14C]isobutyrate. Biochem J 268: 799 –
Sci 61: 9 –19, 1994. 802, 1990.
205. SHENNAN DB AND MCNEILLIE SA. High affinity (Na⫹ Cl⫺)-depen- 224. THOMPSON GE. Cortisol and regulation of tight junctions in the
dent taurine transport by lactating mammary tissue. J Dairy Res mammary gland of the late-pregnant goat. J Dairy Res 63: 305–308,
61: 335–343, 1994. 1996.
206. SHENNAN DB AND MCNEILLIE SA. Milk accumulation down reg- 225. THREADGOLD LC, COORE HG, AND KUHN NJ. Monosaccharide
ulates amino acid uptake via systems A and L by lactating mam- transport into lactating-rat mammary acini. Biochem J 204: 493–
mary tissue. Horm Metab Res 26: 565– 618, 1994. 501, 1982.
207. SHENNAN DB, MCNEILLIE SA, AND CURRAN DE. The effect of a 226. THREADGOLD LC AND KUHN NJ. Monosaccharide transport in the
hyposmotic shock on amino acid efflux from lactating rat mam- mammary gland of the intact lactating rat. Biochem J 218: 213–219,
mary tissue: stimulation of taurine and glycine efflux via a pathway 1984.
distinct from anion exchange and volume-activated anion channels. 227. TODDYWALLA VS, KARI FW, AND NEVILLE MC. Active transport
Exp Physiol 79: 797– 808, 1994. of nitrofurantoin across a mouse mammary epithelial monolayer.
208. SHENNAN DB, MCNEILLIE SA, JAMIESON EA, AND CALVERT DT. J Pharmacol Exp Ther 280: 669 – 676, 1997.
Lysine transport in lactating rat mammary tissue: evidence for an 228. TURNER MD, RENNISON ME, HANDEL SE, WILDE CJ, AND BUR-
interaction between cationic and neutral amino acids. Acta Physiol GOYNE RD. Proteins are secreted by both constitutive and regu-
Scand 151: 461– 466, 1994. lated secretory pathways in lactating mouse mammary epithelial
209. SHENNAN DB, MILLAR ID, AND CALVERT DT. Mammary-tissue cells. J Cell Biol 117: 269 –278, 1992.
amino acid transport systems. Proc Nutr Soc 56: 177–191, 1997. 229. UCHIDA S, KWON HM, YAMAMUCHI A, PRESTON AS, MARUMO
210. SHILLINGFORD JM, CALVERT DT, BEECHEY RB, AND SHENNAN F, AND HANDLER JS. Molecular cloning of the cDNA for an MDCK
DB. Phosphate transport via Na⫹-Pi cotransport and anion ex- cell Na⫹- and Cl⫺-dependent taurine transporter that is regulated
change in lactating rat mammary tissue. Exp Physiol 81: 273–284, by hypertonicity. Proc Natl Acad Sci USA 89: 8230 – 8234, 1992.
1996. 230. ULLRICH KJ, RUMRICH JG, DAVID C, AND FRITZSCH G. Biosub-
211. SHILLINGFORD JM, WOOD JS, SHENNAN DB, SHIRAZI- strates: substances that interact with both renal contraluminal
BEECHEY SP, AND BEECHEY RB. Determination of the sequence organic anion and organic cation transport systems. Pflügers Arch
of a mRNA from lactating sheep mammary gland that encodes a 425: 300 –312, 1993.
protein identical to the Na⫹-dependent glucose transporter 231. VERMA N AND KANSAL VK. Characterisation of the routes of
(SGLT1) (Abstract). Biochem Soc Trans 25: 467S, 1997. methionine transport in mouse mammary glands. Indian J Med Res
212. SINGER KL, STEVENSON BR, WOO PL, AND FIRESTONE GL. 98: 297–304, 1993.
Relationship of serine/threonine phosphorylation/dephosphoryla- 232. VERMA N AND KANSAL VK. Characterisation and starvation in-
tion signaling to glucocorticoid regulation of tight junction perme- duced regulation of methionine uptake sites in mouse mammary
ability and ZO-1 distribution in non-transformed mammary epithe- gland. Indian J Exp Biol 33: 516 –520, 1995.
lial cells. J Biol Chem 269: 16108 –16115, 1994. 233. VINA J, PUERTES IR, ESTRELA JM, VINA JR, AND GLABIS JL.
213. SMITH KM, AND SHENNAN DB. K⫹ (Rb⫹) transport by a mammary Involvement of ␥-glutamyltransferase in amino-acid uptake by the
secretory cell apical membrane fraction isolated from goats’ milk. lactating mammary gland of the rat. Biochem J 194: 99 –102, 1981.
Exp Physiol 75: 349 –358, 1990. 234. VINA J, PUERTES IR, SAEZ GT, AND VINA JR. Role of prolactin in
214. SMITH-KIRWIN SM, O’CONNOR DM, JOHNSTON J, DE LANCEY amino acid uptake by the lactating mammary gland of the rat. FEBS
E, HASSINK SG, AND FUNANAGE VL. Leptin expression in human Lett 126: 250 –252, 1981.
mammary epithelial cells in breast milk. J Clin Endocrinol Metab 235. VINA JR, PALACIN M, PUERTES IR, HERNANDEZ R, AND VINA J.
83: 1810 –1813, 1998. Role of the ␥-glutamyl cycle in the regulation of amino acid trans-
215. SPITZWEG C, JOBA W, EISENMENGER W, AND HEUFELDER AE. location. Am J Physiol Endocrinol Metab 257: E916 –E922, 1989.
Analysis of human sodium iodide symporter gene expression in 236. VINA JR, PUERTES IR, MONTORO JB, RODRIGUEZ A, AND VINA
extrathyroidal tissues and cloning of its complementary DNA from J. Are the ␥-glutamyl-amino acids signals for the amino acid uptake
salivary gland, mammary gland, and gastric mucosa. J Clin Endo- by lactating mammary gland? Biochem Soc Trans 14: 311–312,
crinol Metab 83: 1746 –1751, 1998. 1986.
216. STELWAGEN K, FARR VC, DAVIS SR, AND PROSSER CG. EGTA- 237. VINA JR, PUERTES IR, MONTORO JB, AND VINA J. Effect of
induced disruption of epithelial cell tight junctions in the lactating starvation and refeeding on amino acid uptake by mammary gland
caprine mammary gland. Am J Physiol Regulatory Integrative of the lactating rat. Biochem J 216: 343–347, 1983.
Comp Physiol 269: R848 –R855, 1995. 238. VINA JR, PUERTES IR, PALACIN M, RODRIGUEZ A, AND VINA J.
217. STELWAGEN K, FARR VC, MCFADDEN HA, PROSSER CG, AND Role of oxoproline in amino acid transport in placenta and lactat-
DAVIS SR. Time course of milk accumulation-induced opening of ing mammary gland. Biochem Soc Trans 14: 1056 –1057, 1986.
mammary tight junctions and blood clearance of milk components. 239. VINA JR, PUERTES IR, RODRIGUEZ A, SAEZ GT, AND VINA J.

Effect of fasting on amino acid metabolism by lactating mammary 253. WOODING FBP. Formation of the milk fat globule membrane
gland: studies in women and rats. J Nutr 117: 533–538, 1987. without participation of the plasmalemma. J Cell Sci 13: 221–235,
240. VINA JR, PUERTES IR, AND VINA J. Effect of premature weaning 1973.
on amino acid uptake by the mammary gland of lactating rats. 254. WOODING FBP, PEAKER M, AND LINZELL JL. Theories of milk
Biochem J 200: 705–708, 1981. secretion: evidence from the electron microscopic examination of
241. VIRK SS, KIRK CJ, AND SHEARS SB. Ca2⫹ transport and Ca2⫹- milk. Nature 226: 762–764, 1970.
dependent ATP hydrolysis by Golgi vesicles from lactating rat 255. YUDILEVICH DL AND MANN GE. Unidirectional uptake of sub-
mammary glands. Biochem J 226: 741–748, 1985. strates at the blood side of secretory epithelia: stomach, salivary
242. VREESWIJK JHA, DE PONT JJHHM, AND BONTING SL. Occur- gland, pancreas. Federation Proc 41: 3045–3053, 1982.
rence and properties of (Na⫹-K⫹)ATPase in immature, lactating 256. ZAMMIT VA. Mechanisms of regulation of the partition of fatty
and involuted guinea pig mammary gland. Biochim Biophys Acta acids between oxidation and esterification in the liver. Prog Lipid
330: 173–185, 1973.
Res 23: 39 – 68, 1984.
243. WANG S, WEBB KE, AND AKERS MR. Peptide-bound methionine
257. ZAMMIT VA, GRANT A, RAMSAY RR, BURNS C, AND SHENNAN
can be a source of methionine for the synthesis of secreted proteins
by mammary tissue explants from lactating mice. J Nutr 126: DB. Expression of a sodium-dependent L-carnitine transporter in
1662–1672, 1996. lactating rat mammary tissue. Biochem Soc Trans 26: 96S, 1998.
244. WENDRINSKA A, ADDEY CVP, ORANGE PR, BODDY LM, HEN- 258. ZEISEL SH. Dietary choline: biochemistry, physiology and pharma-
DRY KAK, AND WILDE CJ. Effect of a milk fat globule membrane cology. Annu Rev Nutr 1: 95–121, 1981.
fraction on cultured mouse mammary cells (Abstract). Biochem 259. ZEISEL SH, MAR MH, ZHOU Z, AND DA COSTA KA. Pregnancy and
Soc Trans 21: 220S, 1993. lactation are associated with diminished concentrations of choline
245. WEST DW. Energy-dependent calcium sequestration activity in a and its metabolites. J Nutr 125: 3049 –3054, 1995.
Golgi apparatus fraction derived from lactating rat mammary 260. ZERANGUE N AND KAVANAUGH MP. Interaction of L-cysteine
gland. Biochim Biophys Acta 673: 374 –386, 1981. with a human excitatory amino acid transporter. J Physiol (Lond)
246. WHITE MD, KUHN NJ, AND WARD S. Permeability of lactating-rat 493: 419 – 423, 1996.
mammary gland Golgi membranes to monosaccharides. Biochem J 261. ZETTL KS, SJAASTAD MD, RISKIN PM, MACHEN TE, AND FIRE-
190: 621– 624, 1980. STONE GL. Glucocorticoid-induced formation of tight junctions in
247. WILDE CJ, ADDEY CVP, BODDY LM, AND PEAKER M. Autocrine mouse mammary epithelial cells in vitro. Proc Natl Acad Sci USA
regulation of milk secretion by a protein in milk. Biochem J 305: 89: 9069 –9073, 1992.
51–58, 1995. 262. ZHOA F-Q, DIXON WT, AND KENNELLY JJ. Localization and gene
248. WILDE CJ, ADDEY CVP, BODDY-FINCH LM, AND PEAKER M. expression of glucose transporters in bovine mammary gland.
Autocrine control of milk secretion: from concept to application. Comp Biochem Physiol B Biochem 115: 127–134, 1996.
In: Intercellular Signalling in the Mammary Gland Proceedings of 263. ZHOA F-Q, GLIMM DR, AND KENNELLY JJ. Distribution of mam-
the Hannah Symposium, Ayr, 13–15 April 1994, edited by C. J. malian facilitative glucose transporter messenger RNA in bovine
Wilde, M. Peaker, and C. H. Knight. New York: Plenum, 1995, p. tissues. Int J Biochem 25: 1897–1903, 1993.
227–237. 264. ZHOA F-Q, MOSELY WM, TUCKER HA, AND KENNELLY JJ. Regu-
249. WILDE CJ, ADDEY CVP, BRYSON JM, FINCH LMB, KNIGHT CH,
lation of glucose transporter gene expression in mammary gland,
AND PEAKER M. Autocrine regulation of milk secretion. In: Mam-
muscle, and fat of lactating cows by administration of bovine
mary Development and Cancer, edited by P. S. Rudland, D. G.
growth hormone and bovine growth hormone-releasing factor. J
Fernig, S. Leinster, and G. G. Lunt. London: Portland, 1997, p.
81–90. Anim Sci 74: 183–189, 1996.
250. WILDE CJ, PEAKER M, AND KNIGHT CH. Intercellular signalling in 265. ZHOA F-Q, OKINE EK, SUOMINEN AH, GLIMM DR, AND KEN-
the mammary gland. In: Proceedings of the Hannah Symposium, NELLY JJ. Expression of Na⫹/glucose cotransporter gene in the
Ayr, 13–15 April 1994. New York: Plenum, 1995. gastrointestinal tract and the mammary gland of lactating cows
251. WILDE CJ, PEAKER M, AND TAYLOR E. Biological Signalling and (Abstract). J Dairy Sci. 79 Suppl: 145, 1996.
the Mammary Gland. Ayr, Scotland: Hannah Research Institute, 266. ZIV G AND SULMAN FG. Effects of probenecid on the distribution,
1997. elimination, and passage into milk of benzylpenicillin, ampicillin
252. WOO PL, CHA HH, SINGER KL, AND FIRESTONE GL. Antagonistic and cloxacillin. Arch Int Pharmacodyn Ther 207: 373–382, 1974.
regulation of tight junction dynamics by glucocorticoids and trans- 267. ZULAK IM AND KEENAN TW. Citrate accumulation by a Golgi
forming growth factor-beta in mouse mammary epithelial cells. apparatus-rich fraction from lactating bovine mammary gland. Int
J Biol Chem 271: 404 – 412, 1996. J Biochem 15: 747–750, 1983.

Transport of Milk Constituents by The Mammary Gland - Sheenan

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PHYSIOLOGICAL REVIEWS

Vol. 80, No. 3, July 2000

Hannah Research Institute, Ayr, Scotland

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I. INTRODUCTION various mammary gland preparations, effects on trans-

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The perfused lactating mammary gland has been suc-

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FIG. 2. Diagram showing the known major

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bursting the taut epithelium (69, 161). Similarly, in the

IV. SODIUM, POTASSIUM, AND CHLORIDE

The mammary gland is able to generate and maintain

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C. Effects of Prolactin on Naⴙ, Kⴙ, and

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calcium present in milk (144). Neville and Peaker (146)

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VIII. CITRATE TRANSPORT X. STRETCH-SENSITIVE ION TRANSPORT

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TABLE 1. Kinetic properties of mammary tissue amino acid transport systems

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been suggested that amino acids derived from circulating

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include glutathione (␥-glutamyl-cysteinylglycine) and cer-

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