BIOLOGY OF REPRODUCTION 38, 3 1-38 (1988)

The Biochemistry of Female Reproductive Tissues Studied by

31Phosphorus Nuclear Magnetic Resonance Spectroscopy:
Effects of Pregnancy, Hormonal Manipulation, and Disease’

M. J. DAWSON,2 D. K. McFARLANE,
B. L. McFARL!N, E. A. NOYSZEWSKI, and S. R. TRUPIN

Departments of

Downloaded from https://academic.oup.com/biolreprod/article-abstract/38/1/31/2763481 by guest on 04 March 2019

Physiology and Biophysics and Obstetrics and Gynecology
University of Illinois at Urbana-Champaign
Urbana, Illinois 61801

INTRODUCTION and diseased female reproductive tissues, 2) to make

Studies of the bioenergetics of normal and ab- apparent some of the potentials of NMRS techniques
normal reproductive tissues have been limited by the that are already available or are in the process of
difficulties that accompany application of traditional development for application in studies of reproduc-
analytical techniques. Over the past. dozen years, tive biology, and 3) to evaluate the potential of
nuclear magnetic resonance spectroscopy (NMRS) has NMRS for diagnosis, prognosis, and assessment of
opened unparalleled opportunities for noninvasive treatment for diseases of female reproductive tissue.
studies of function and metabolism in cell suspensions Additional information on each of these topics is
and isolated perfused tissues as well as in selected available in a recent monograph on magnetic resonance
volumes within animals, normal human subjects, and imaging and spectroscopy of the reproductive system
patients. Localized spectroscopic studies using 3,P, (McCarthy and Haseltine, 1987).
‘3C, and ‘H are now being used to observe regional
differences in chemical composition as well as varia- The Technique
tions in tissue metabolism and enzyme regulation. NMRS techniques rely upon the excitation and
Other nuclei are used to monitor ion transport and
detection of an electromagnetic resonance signal in
distribution within a number of organs.
the radio-frequency range. Such signals are emitted
Applications of these techniques to reproductive by certain magnetically susceptible atomic nuclei
tissues has been relatively slow, with progress impeded
when a sample containing them (which may be as
by the low concentrations of metabolites within the simple as a solution in a test tube or as complex as
tissue volumes of interest, and by the difficulty of the human body) is placed in a strong magnetic field
sampling reproductive tissue sites noninvasively.
and excited by a pulse of electromagnetic energy of
However, a number of laboratories, including our
appropriate frequency. The resonance frequency of
own, has obtained useful results in studies of isolated
the emitted radiation is characteristic of the atomic
reproductive tissues and cell suspensions, and
nucleus and the nature of its immediate chemical
theoretical calculations show that meaningful non-
environment.
invasive NMRS studies of reproductive tissues in Figure 1A shows the first published NMR spectrum
human subjects are feasible. of a reproductive tissue (Dawson and Wray, 1983).
The present paper has three goals: 1) to summarize This is a 31P NMR spectrum of four nonpregnant uteri
the work from our own and other laboratories using isolated from rat. The tissues were superfused at 18#{176}C
NMRS to study function and metabolism in normal with 25 mM bicarbonate Kreb’s solution bubbled
with 95% 02 /5% CO2. The resonances are identified
on the basis of their position on the frequency (X)
‘We are grateful to the Research Board of the University of Illinois axis after appropriate studies of model solutions to
at Urbana-Champaign and to the Servants United Foundation for support
determine the resonance frequency of biological
of this work.
2 Reprint requests. compounds under physiological conditions.

31
32 DAWSON ET AL.

The peak at -9.6 p.p.m., between the 3 and a

PCr
peaks, is in the position of uridine diphosphoglucose
(UDPG). Peaks from phosphocreatine (PCr) and
inorganic phosphate (Pi) can be seen clearly and
unambiguously assigned. The position of the Pi peak
is sensitive to pH, and in this spectrum, two distinct
peaks are observed: the intracellular Pi is on the
right, representing pH 7.0, and the peak on left is
from Pi in the bathing medium at pH 7.4. The peak

Downloaded from https://academic.oup.com/biolreprod/article-abstract/38/1/31/2763481 by guest on 04 March 2019

at 6.8 p.p.m., in the phosphomonoester (PME) region
of the spectrum, is in the position of phospho-
ethanol-amine, and the shoulder on the right of this
peak may represent phosphocholine. Both of these
compounds have been shown to be present in myo-
metrium (Harkness and Harkness, 1954). There are
three very small peaks apparent in the phosphodiester
L I I
region (PDE) of the spectrum, at 2.3, 3.1, and 3.6
I0 0 -10 20 p.p.m. Two of these have been identified by Degani
Chemical shift (ppm) (1987) in mammalian female reproductive tissues as
FIG. 1. 31P Nuclear magnetic resonance spectrum of four isolated glycerol-phosphorylethanolamine (3.5 p.p .m.) and
mature and nonpregnant rat uteri obtained at 81 MHz. The X-axis is glycerol-phosphorylcholine (3.1 p.p.m.).
frequency in parts per million (p.p.m.) difference from the phospho-
creatine (PCr)-reference frequency. The V-axis is signal intensity. This
The first noninvasive spectrum of reproductive
is the accumulated result of 1198pulsesof30.isdurationat2-sintervals. tissue was recently obtained from a localized volume
Uteri were superfused at 18#{176}Cwith 25 mM bicarbonate Kreb’s solution
bubbled with 95% 02/5% CO2. See text for peak assignments. (From
within the uterus of an anesthetized rabbit (c.f.
Dawson and Wray, 1983). Haseltine, 1987); the animal was placed in a 33-cm
horizontal-bore magnet operating at 4.7 Tesla. Such
localized studies are made possible by two alterations
from the traditional design of NMR spectrometers:
1) The magnet is positioned horizontally and is
In Figure 1A, the peaks at -16.5, -7.5, and -2.6
parts per million (p.p.m.) are from nucleotide phos- large enough to admit a small animal. Magnets having
pliate (NP). The individual nucleotides resonate so an internal diameter of 60 cm, large enough to
accommodate an adult human, are also available. For
closely to each other that they are indistinguishable
in the spectra. Adenine is the major (75%) nucleotide the present, they are lower in magnetic field strength
in the uterus, uridine and guanidine each contributing and therefore in sensitivity than the most advanced
around 10% (Oliver and Kellie, 1970). The peak at animal systems.
2) The magnetic field is shaped so that signals are
-16.5 p.p.m. arises from the middle (j3) phosphorus,
received from a localized sensitive volume. In the
and is therefore composed solely of triphosphate
(NTP). Its position indicates that approximately half simplest case, the shaping of the field is done by
placing a “surface coil” in contact with the tissue of
of the total NTP is in the form of MgNTP2. The
other two peaks represent phosphorus at the ribose interest, e.g. within one horn of the uterus. This
(a) end and the terminal (y) end of the molecule, and small, circular, radio-frequency coil is used to apply
therefore contain contributions from both di- and the excitation pulses and detect the resulting signals.
triphosphates. The concentrations of diphosphates A number of more sophisticated and versatile localiza-
(NDP) can be obtained by subtraction of the j3- tion methods are now available, and new ones are
rapidly evolving.
from the ‘v-peak intensity. Nicotinamide adenine
dinucleotide (NAD) and NAD phosphate (both their
oxidized and reduced forms) resonate as a shoulder to NMR-Sensitive Nuclei for Studies of Living Tissues
the a-peak; therefore, the total concentration of these The usefulness of a particular nucleus in extracting
pyridine nucleotides (PN) can be obtained by biological and metabolic information depends both
subtraction of the 7NP from the aNP peak intensities. upon the strength of the NMR signal emitted and
 NMRS OF FEMALE REPRODUCTIVE TISSUES 33
upon its natural abundance in the body (or the ability free Mg2 (Gupta and Gupta, 1987).
to introduce it into the body). Some useful nuclei 13C Natural abundance and enrichment. ‘3C is
that either have been or are likely soon to be used in present in all organic compounds. However, having a
studies of reproductive biology include ‘H, ‘9F, 31P, natural abundance of only 1.1%, it yields very weak
and ‘3C. signals. The only resonance peaks normally observed
31P Studies of bioenergetics. Owing to the impor- in a ‘3C spectrum from mammalian tissues arise from
tance of phosphorus compounds in cellular energetics, the large amounts of carbon present in fatty tissues,
the naturally occurring nucleus, 3’P (99% isotopic membranes, and glycogen. Natural abundance 13C
abundance), has featured predominantly in NMRS of NMR is now being used to monitor physiological and

Downloaded from https://academic.oup.com/biolreprod/article-abstract/38/1/31/2763481 by guest on 04 March 2019

living tissues. The 3’P nucleus gives strong signals pathological changes in glycogen content in such
from a small number of important compounds tissues as liver and skeletal muscle. The usefulness of
present in living systems and also yields an estimate natural abundance ‘3C NMRS for studies of reproduc-
of intracellular pH and free Mg2. Thus, in Figures 1, tive tissues is limited by the long period (often
2, 4, and 5 of this paper, the intensities of the peaks several hours) required to obtain a meaningful
attributed to NTP, NP, PN, PCR (when it occurs), spectrum.
and Pi can yield a direct assessment of the energetic The power of ‘3C NMRS can be greatly extended
status of an organ. This type of information can then by introducing specific ‘3C-enriched compounds in a
be correlated with tissue function under a number of manner analogous to more familiar experiments using
normal physiological and pathological conditions. the radioactive ‘4C isotope. In addition to being
Energy is required not just for activity but also for noninvasive and nondestructive, ‘3C NMRS provides
growth, and 31P-containing metabolites are involved in the ability to compare the site of the original ‘3C
this process as well. The PMEs and PDEs that have so label with its sites in metabolic products. The precise
far been identified in mammalian tissue are all inter- resonance position of a ‘3C-labeled carbon is dependent
mediates in the synthesis or degradation of membrane on whether its nearest neighbors are ‘2C or ‘3C.
phospholipids. It has been suggested that the concen- Thus, the ‘3C label may be split due to the presence
trations of these substances are related to regulation of ‘3C at a neighboring position. The extent to which
of membrane composition (Burt, 1985). The finding the splitting occurs in the spectrum of a product
that these peaks, especially PME, are large in most compound indicates the extent to which reactants
cancers studied, and may vary with the state of the derived from the initial labeled substrate combined to
tumor (Evanochko et al., 1984), tends to focus a form the product. This, in turn, indicates to what
great deal of attention upon them. extent nonlabeled pathways are involved. Applica-
The precise resonance positions of charged species tions of ‘3C NMR to the study of metabolic regula-
are dependent on their chemical environment. For tion in the living cell have recently been reviewed
example, Pi is found within cells almost entirely as (Cohen, 1987).
H2P04 1 and HPO4 2 and the pK that determines 1H NMRS with water suppression. Although ‘H is
the relative proportions of these two species is close the most sensitive nucleus for NMR spectroscopy, its
to physiological pH. The two ionic forms of Pi have use in metabolic studies has been inhibited by the
slightly different resonance frequencies, and a single formidable problem of observing signals from milli-
Pi peak appears in an intermediate position depending molar concentrations of metabolites in the presence
upon relative concentrations. Thus, after appropriate of 50 M H20. Large signals from storage fats also
calibration, the position of the Pi peak can be used to cause difficulties for studies of most mammalian
determine intracellular pH. An analogous situation tissues. One of the recent methodological develop-
pertains to NTP. After appropriate calibration, ments of great importance in spectroscopy of living
the position of the j3NTP peak can be used to deter- tissue has been the application of water and lipid
mine intracellular free Mg2. Measurements of con- suppression techniques, which allows observation of
centrations of ionic species of Pi and NTP are useful the ‘H spectrum of metabolites down to 0.1 mmol
in themselves, and they yield absolute concentrations kg’ in concentration. A number of closely related
of both H and Mg24, which influence most cellular techniques are now being applied to studies of living
activities. Reviews are available on the use of NMRS tissues, and the methodologies and results have
to measure intracellular pH (Gullies et al., 1982). and recently been reviewed (Avison et al., 1986; Redfield,
34 DAWSON ET AL.

1986). Of potentially great importance in studies of Studies of the effects of hormonal interventions on the
reproductive tissues is the fact that changes in lactate 31P NMR spectrum of the myometrium have been
as a result of exertion or ischemia are both discernable carried out by Degani and coworkers (1984). Compari-
and quantifiable using ‘H NMRS. son of the bioenergetic properties of the uterus with
Intra- to extracellular Na and K gradients. those of other smooth muscles have been carried out
Chemical shift reagents of the lanthanide series offer by other laboratories (Hellstrand and Vogel, 1985;
great promise in the study of compartmentation of Kushmerick et al., 1986).
NA and K. When introduced into an extracellular Figure 2 shows spectra from nonpregnant (above)
fluid compartment, they shift the resonance position and term pregnant (below) isolated human uterus,

Downloaded from https://academic.oup.com/biolreprod/article-abstract/38/1/31/2763481 by guest on 04 March 2019

of the ions to which they bind away from that studied under identical conditions. The spectrum
of the remaining intracellular ions. They have been from term-pregnant myometrium shows elevated
used to distinguish intra- from extracellular Na and concentrations of PCr, UDPG, and PDE; these changes
K signals in cell suspensions, isolated hearts, and were all statistically significant at p<0.05 or better
skeletal muscle of anesthetized animals (see review by when all our results on term-pregnant (n=12) and
Springer, 1987). Rubidium is also shifted by these nonpregnant (n=7) myometrium were compared. The
reagents, and therefore 85Rb NMR can be used to mean intracellular pH was slightly more alkaline in
monitor the rate of Rb uptake in studies of the term-pregnant than in nonpregnant human myo-
mechanism of ion transport. metrium (7.29 ± 0.01 SEM, n=7 and 7.14 ± 0.01
19F NMR. ‘9F yields strong NMR signals. Coupled SEM, n=12, at 37#{176}C,respectively). There was a
with this is the fact that there is no background signal significant change (p<0.05) as a result of pregnancy
from endogenous compounds. Present applications in the chemical shift difference between the a and p
include observation of the metabolic fate and distribu- NP peaks, indicating that Mg2-binding to NTP
tion of fluorinated anesthetics and anti-cancer drugs. increased and myometrial-free Mg2 increased from
Fluorinated chelators of mono- and divalent cations 0.32 (SEM = 0.005, n=12) to 0.42 mM (SEM = 0.02,
are used as “shift reagents” for determining intra- n=7) as a result of pregnancy.
cellular cation concentrations. These may be func- In both the human and the rat study, PCr was
tionally and chemically similar to fluorescence almost doubled in the pregnant compared to non-
probes. For example, symmetrically substituted pregnant myometrium. This finding is in accord with
difluoro derivatives of 1 ,2-bis (0-aminophenoxy) previous chemical estimates of changes in PCr in
ethane-N’,N’,N’-tetracetic acid; (FBAPTA) show large human uterus as a result of pregnancy (22 5%
‘9F NMRS chemical shifts upon chelation of divalent larger, Cretius, 1957). Since PCr is present in the
cations. After introduction of the nonionized myometrium, as it is in other tissue types, as a
compound into the intracellular compartment and its buffer for the supply of adenosine triphosphate
enzymatic cleavage and entrapment, 5FBAPTA and (ATP), this biochemical change tends to prepare the
similar compounds can be used to measure intra- uterus for the sustained and powerful contractions
cellular calcium concentrations in isolated cells and required to expel the young. UDPG, which is involved
isolated tissues as first described by Smith et al. in the synthesis of glycogen from glucose, is higher in
(1983). concentration in pregnant than in nonpregnant
human myometrium. This could be related to the
increase in glycogen content during pregnancy, and to
NMRS Studies of Reproductive Tissues
the reliance of the myometrium on glycogen, rather
Myometrium. A large number of biochemical and than on blood-glucose, to fuel contractions (Chew
structural changes are known to occur in the uterus and Rinard, 1979). The changes in pH and Mg2 as a
to support pregnancy, and these are rapidly reversed result of pregnancy could have profound functional
during the period of involution that follows delivery significance in terms of both energy metabolism and
of the fetus (Reynolds, 1949). We have found that the activation of contractions.
NMRS can be used to investigate the bioenergetic The biochemical and structural changes that occur
changes that accompany pregnancy, parturition, during pregnancy must be rapidly reversed during the
and involution in the isolated rat uterus (Dawson and period of involution. We have therefore investigated
Wray, 1985) and in myometrium obtained from human the biochemical changes that follow delivery of the
subjects (Noyszewski et al., unpublished results). fetuses. Figure 3 shows the changes in concentrations
 NMRS OF FEMALE REPRODUCTIVE TISSUES 35
Non- pregnant mean ± se
PME
4.4 P pregnant

NP, non-pregnant
3.6

2.8

2.0...
+PME

Downloaded from https://academic.oup.com/biolreprod/article-abstract/38/1/31/2763481 by guest on 04 March 2019

: 1.21-

NIP
. 101 F-
0.8.

0.61
Pregnant
P1
0.4L
1.4 8

1.2

1.oL\IL5

0.8
II I I I

P0 2 4 6 21 NP
day post-partum

FIG. 3. The postpartum changes in concentration (relative to

nucleotide triphosphate, that did not change with time) of phospho-
monoesters (PME), inorganic phosphate (Pi), and phosphocreatine

10
- I

5
I

0 -5 -10 -15 - -25

(PCr)
next
errors
in the isolated
to the points
indicated
rat uterus.
on the PCr curve.
by vertical lines.
The numbers
Values
P = term-pregnant,
of observations
are means
are given
with standard
NP = nonpregnant.
Chemical Shift (ppm) (From Dawson and Wray, 1985).

FIG. 2. 31P Nuclear magnetic resonance spectra of nonpregnant

(obtalned at hysterectomy) and term-pregnant (obtained from the edge
of a Cesarean incision) human myometrium run at 121 MHz using a
pulse duration of 20 ss (tip angle, 350) and pulse interval of 1.5 S. The
V-axes have been adjusted so that for any given peak, equal signal Figure 3, there was a significant intracellular acidifica-
intensities in the two spectra represent equal concentrations of
metabolite. Samples were placed in ice-cold oxygenated de-Jalon’s
tion, which had a time course similar to the changes
solution immediately upon clinical inspection (within 1 to 2 mm of in the PME-peak intensity.
removal) and transported to the spectrometer where they were super-
To determine whether these changes in metabolite
fused with oxygenated de-Jalon’s solution throughout the experiment.
Samples were obtained with written consent of the patients and with concentrations result from mechanical activity during
the approval of the Institutional Review Board of the University of parturition, experiments on Cesarean-sectioned
Illinois, Urbana-Champaign. See text for peak assignments.
animals were undertaken. Although Cesarean-
sectioned uteri undergo far less muscular work than
do normal uteri, we found that the operatiOn had no
of phosphorus metabolites that occur after parturi- effect on phosphorus metabolite concentrations or
tion in the rat. The PCr fell by one-third after intracellular pH. The results therefore do not support
delivery, and the Pi doubled. The PME almost doubled the hypothesis that smooth muscle activity is
in size during the first day postpartum, and then responsible for the metabolic changes observed.
decreased so that on Day 2 it was only 20% greater Instead, they are consistent with the hypothesis that
than in nonpregnant animals. Although not shown in uterine metabolism is under hormonal control.
36 DAWSON ET AL.

obtained, suggests that 3’P NMRS could become a

method for early diagnosis of this disease.
Spectroscopy of the placenta. Figure 5 shows
typical 3’P NMR spectra from placentae of first,
Pu second, and third trimester gestational age. With the
exception of PCr, which is absent in most spectra of
placentae, the same peaks appear as are seen in
uterine tissue. There is a clear decrease in the size of
the PME peak and an increase in the Pi peak as the

Downloaded from https://academic.oup.com/biolreprod/article-abstract/38/1/31/2763481 by guest on 04 March 2019

placenta matures. There was a significant decrease in

First Trimester
0 -5 -10 -15 -20 PPM PME NP

FIG. 4. 31P Nuclear magnetic resonance spectrum of nonpregnant

myometrium obtained at hysterectomy from an 87-year-old patient
with endometrial adenocarcinoma, which did not appear to the pathol-
ogist to invade the myometrium. Data accumulation and processing
were the same as for Figure 2. See text for peak assignments.

The change in PME peak requires particular notice.

10 5 0 -5 -tO -IS -20 -25 PPM
Phosphoethanolamine, which is probably the main
contributor to this peak, is a product of lipid Second Trimester
catabolism. The increase in size of the PME peak seen
after parturition may well be related to the decrease
in cell size, and thus membrane area, occurring during
uterine involution. The time courses of the changes
coincide; the rat uterus halves in size during the first
24-36 h postpartum (Harkness and Harkness, 1954),
and the largest changes in the PME peak are seen
during the same period. -5 -10 -15 -20 -25 PPM
Hormonal control of myometrial metabolism: Third Trimester
reproductive tissue tumors. Studies of breast tumors
show that benign breast tumors have spectral charac-
teristics that are quite different from neoplasms
(Degani et al., 1986), and that the hormonal response
of breast carcinoma can be predicted from the 31P
spectrum (Victor et al., 1987). We have studied tissue
samples from two patients with endometrial adeno-
carcinoma. Although the myometrial tissue obtained
appeared to be normal at a macroscopic level, the 31P
NMR spectra, one of which is shown in Figure 4,
showed a significantly raised PME (159%, p<0.OO1)
and increased Pi (192%, p<0.001) as compared to the
pooled results on all other nonpregnant uteri. No PCr
10 5 0 -5 -10 -15 -20 -25 PPM
peak was visible. These same spectral characteristics
FIG. 5. 31P Nuclear magnetic resonance spectra of first (11 weeks),
have been observed in cultured human adeno- second (21 weeks) and third (39 weeks) trimester human placentae
carcinoma cells (Fantini et al., 1987). The ability to superfused with oxygenated de-Jalon’s solution at 4#{176}C.The V-axes
have been adjusted so that for any given peak, equal signal intensities
deduce the presence of adenocarcinoma from the among the spectra represent equal concentrations of metabolites. See
spectrum, but not the appearance of the tissue text for peak assignments.
 NMRS OF FEMALE REPRODUCTIVE TISSUES 37
PME signal is highest when the tissue is undergoing its
most rapid relative growth is consistent with the
suggestion discussed above that this peak is related to
membrane turnover.
Abnormalities of pregnancy. Both diabetes and
hypertension can lead to structural abnormalities of
the placenta, which could well be associated with
alterations in intracellular milieu and/or in percentages
of various tissue types. We have studied placentae from

Downloaded from https://academic.oup.com/biolreprod/article-abstract/38/1/31/2763481 by guest on 04 March 2019

three hypertensive women; all showed lower PN
(55%, p<O.OOl; Fig. 7) and lower free Mg2 (68%,
p<0.02; Fig. 6) than the normal term placentae. A
sample of uterus was available from one of these
women, and it showed a strikingly higher PN than
L the normal myometrial samples studied (163%,
p<0.001; Fig. 7). The single placental samples ob-
tained from a woman with gestation diabetes had an
elevated PN (162%, <O.OOl; Fig. 7). The patient

Normal Hyper. Diabetic Cocaine

FIG. 6. Free intracellular MgS+ obtained from the relationship

-b -15.86
pMg= 4.11 +Iog
19.02 + 5
55

where b is the chemical shift of the 3-nicotinamide triphosphate peak

,
relative to phosphocreatine. The height of the bars represents the mean
of 6 normal first and 5 normal third trimester placentae, 3 placentae
from hypertensive patients, 1 from a diabetic patient, and 1 from a
cocaine abuser. Standard error bars are given where possible. p values
represent the difference between normal first and third trimester
55
placentae and between normal and hypertensive term placentae.

apparent PN and UDPG and an increase in Mg2 in

third trimester as compared to first trimester placenta.
At present, very little can be said about either the
mechanism or the consequences of these changes. The
placenta is a very heterogeneous tissue, and its
composition changes as gestation continues. The Normal Hyper. Diabetic Cocaine Normal. Hyper.
increase in N content of placental tissue with matura-
FIG. 7. Peak intensities for total pyridine nucleotides, obtained by
tion could be related to the increased Pi transport to subtraction of the a from the ‘y-NP peaks as described in the text.
the fetus, since two-thirds of the total body Pi in Placental values are from the same studies that are shown in Figure 6.
Uterine values represent the pooled results of 14 nonpregnant and 8
term neonates is deposited during the third trimester. pregnant myometria and a single value obtained from the myometriuni
The observation that the intensity of the placental of a hypertensive patient.
38 DAWSON ET AL.

who so far has shown the most substantial changes in regulation in the living cell. In: Gupta RK (ed.), NMR Spectroscopy
of Cells and Organisms, Vol. 1. Boca Raton, FL: CRC Press, pp.
biochemistry was a cocaine user: at the 5% level there 3 1-50
was higher Pi and lower PN and UDPG. Cretius K, 1957. Der kreatin phosphat gehalt menschlicher skelett-und
uterus muskulatur. Geburtshilfe und Gynakologie 149:113-22
At present, we are unable to suggest either the Dawson MJ, Wray S, 1983. 31P nuclear magnetic resonance (NMR)
mechanism or the functional significance of these studies of isolated rat uterus. J Physiol 336: 19-20P
Dawson MJ, Wray 5, 1985. The effect of pregnancy and parturition on
changes. However, it is interesting to relate the
phosphorus metabolites in the rat uterus studied by 31P nuclear
biochemical abnormalities observed in the placentae magnetic resonance. J Physiol 368:19-31
from these patients to the changes observed during Degani H, 1987. NMR spectroscopy studies of the reproductive organs
and associated malignancies. In: McCarthy S, Haseltine F (eds.),
normal placental maturation; all of the differences Magnetic Resonance of the Reproductive System. Thorofare, NJ:

Downloaded from https://academic.oup.com/biolreprod/article-abstract/38/1/31/2763481 by guest on 04 March 2019

observed between normal and abnormal term placen- Slack, Inc., pp. 81-96
Degani H, Horowitz A, Itzchak Y, 1986. Breast tumors: evaluation with
tae relate to substances that change significantly in P-3 1 MR spectroscopy. Radiology 161:53-55
concentration during placental growth. It may be Degani H, Shaer A, Victor TA, Kaye AM, 1984. Estrogen-induced
changes in high-energy phosphate metabolism in rat uterus.
entirely fortuitous, but we note that these possible NMR studies. Biochemistry 23:2572-77
alterations in reducing power (NADP) correlate with Evanochko WT, Ng TC, Glickson JD, 1984. Application of in vivo NMR
spectroscopy to cancer. Magn Reson Med 1:508-34
the tendency toward growth retardation of placenta
Fantini J, Galons J-P, Marvaldi J, Cozzone PJ, Canioni P, 1987. Growth
and fetus in hypertensive patients and the tendency of a human colonic adenocarcinoma cell line (HT29) on micro-
carrier beads: metabolic studies by 31phosphorus nuclear magnetic
to macrosomny of placenta and fetus in diabetic
resonance spectroscopy. IntJ Cancer 39:255-60
patients. The abnormally low Mg2 observed in Gillies RJ, Alger JR, den Hollander JA, Shulman RG, 1982. Intra-
hypertensive placenta could be related to abnormal cellular pH measured by NMR: methods and results. In: Nuccitelli
R, Deamer DW (eds.), Intracellular pH. Its Measurement, Regula-
Ca/Mg balance, and could be a part of the explana- tion and Utilization in Cellular Functions. Vol. 15. New York, NY:
tion for the excessive tone in the placental vasculature. Alan R. Liss, Inc., pp. 341-519
Gupta RK, Gupta P, 1987. 31P NMR measurement of intracellular free
magnesium in cells and organisms. In: Gupta RK (ed.), NMR
CONCLUSIONS Spectroscopy of Cells and Organisms, Vol. II. Boca Raton, FL:
CRC Press, pp. 33-44
Our own work, and that from other laboratories, Harkness MLR, Harkness RD, 1954. The collagen content of the

has demonstrated that 31P NMRS techniques can reproductive tract of the rat during pregnancy and lactation. J
Physiol 123:492-500
reveal a great deal about the biochemistry, physiology, Haseltine F, 1987. Introduction to Magnetic Resonance of the Repro-
and pathology of reproductive tissues. The nature, ductive System. McCarthy 5, Haseltine F (eds.). Thorofare, NJ:
Slack, Inc., pp. ix-xiii
mechanism, and consequences of hormonal control of Hellstrand P, Vogel HJ, 1985. Phosphagens and intracellular pH in
bioenergetics in reproductive tissues is being explored. intact rabbit smooth muscled studied by 31P-NMR. Am J Physiol
248:C320-29
We can look forward to application of additional Kushmerick MJ, Dillon PF, Meyer RA, Brown TR, Krisanda JM,
powerful NMRS techniques to these problems. NMRS Sweeney HL, 1986. 31P NMR spectroscopy, chemical analysis and

can be combined with other methodologies to study free MgS+ of rabbit bladder and uterine smooth muscle. J Biol
Chem 261:14420-429
mechanisms at the molecular level and how these Mccarthy S, Haseltine F, 1987. Magnetic Resonance of the Reproduc-
tive System. Thorofare, NJ: Slack, Inc.
operate in intact tissues.
Noyszewski EA, Trupin SR, McFarlin BL, McFarlane DK, Dawson MJ,
1987. 31Phosphorus nuclear magnetic resonance examination of
ACKNOWLEDGMENTS female reproductive tissues. In preparation for Am J Obstetrics
and Gynecology
We thank Dr. Stanley Bobowski, Departmentof Pathology, Burnham Oliver JM, Kellie AE, 1970. The effects of oestradiol on the acid
Hospital, Champaign, Illinois, for his help and advice. This study could soluble nucleotides of rat uterus. Biochem J 119:187-91
not have succeeded without the cooperation of the obstetrical nursing Redfield AG, 1986. Special problems of NMR in H30 solution. In:
staffs at Burnham and Mercy Hospitals. Bradbury EM, Nirolini C (eds.), NMR in the Life Sciences. New
York: Plenum, pp. 1-10
Reynolds SRM, 1949. The Physiology of the Uterus. New York: Hoeber
REFERENCES
Smith GA, Hesketh RT, Metcalfe JC, Feeney J, Morris PG, 1983.
Avison MJ, Hetherington HP, Shulman RG, 1986. Application of NMR Intracellular calcium measurements by ‘#{176}FNMR of fluorine-
to studies of tissue metabolism. Annu Rev Biophys Biophys Chem labeled chelators, PNAS 80:7178-82
15: 3 77-402 Springer CS, 1987. Measurement of metal cation compartmentation in
Burt CT, 1985. Phosphodiesters and NMR: a tale of rabbits and tissue by high-resolution metal cation NMR. Ann Rev Biophys
chickens. TIBS 10:404-06 Biophys Chem 16:376-99
Chew CS, Rinard GA, 1979. Glycogen levels in the rat myometrium at Victor TA, Wiebolt RC, Shattuck MC, Brodin AG, Degani H, 1987.
the end of pregnancy and immediately postpartum. Biol Reprod Prediction of hormonal response of human breast carcinoma by
20:1111-14 31P NMR spectroscopy. In: McCarthy S (ed.), Magnetic Resonance
Cohen SM, 1987. Applications of ‘3C NMR to the study of metabolic of the Reproductive System. Thorofare, NJ: Slack, Inc., pp.67-80