Effect of plasma osmolality on steady-state
fluid shifts in perfused cat skeletal muscle
MARC T. HAMILTON, DIANNE S. WARD,
Departments of Exercise Science and of Physiology,
Columbia, South Carolina 29208
Hamilton, Marc T., Dianne S. Ward, and Philip D.
Watson. Effect of plasma osmolality on steady-state fluid shifts
in perfused cat skeletal muscle. Am. J. Physiol. 265 (Regulatory
Integrative Comp. Physiol. 34): R1318-R1323, 1993.-Fluid re-
distribution in isolatedperfusedcat calf musclecausedby rapid
increasesin plasma osmolality was studied using NaCl or su-
crose.Extracellular tracers (51Cr-labeledEDTA or [3H]manni-
tol) were added to the perfusate 90 min before solutes were
added,and samplesweretaken from plasmaimmediately before
osmolality was increasedand 17,40, and 65 min later. Intersti-
tial fluid volume (IFV) was calculated as extracellular volume
(ECV) minus plasma volume (Evans blue dye). Total tissue
water changes(ATTW) were measuredby continuous recording
of tissue weight. Change in intracellular volume (AICV) was
obtained from ATTW - AIFV. TTW, IFV, ICV, and plasma
osmolality were in steady state after 17 min. Changesin hydro-
static and colloid osmotic pressurewere insignificant in com-
parison with small-moleculeosmotic pressurechanges.The ap-
parent volume of TTW participating in the fluid shift averaged
65 + 1 ml/100 g (SE) over a wide rangeof osmolality increases.
In contrast to the largechangesin TTW, IFV wasnot alteredby
osmolality. Thus decreasesin TTW were similar to cell dehy-
dration. Hence, increasesin plasmavolume induced by hyper-
tonic fluids may comeentirely at the expenseof cell volume, not
interstitial volume.
hyperosmolality; interstitial fluid volume; cell volume; hyper-
tonic solutions; sucrose
ACUTE INCREASES in plasma osmolality, such as occur
during hypertonic fluid resuscitation (14)) exercise (12))
hemorrhage (7, 9), or hyperglycemia (l), cause impor-
tant changes in water and electrolyte homeostasis. Be-
cause steady-state osmolality must be equal in all the
principal body fluid compartments, rapid fluid shifts
from one part of the body must cause either water or
solute changes in the other organs. Thus, for any os-
motic disturbance, skeletal muscle is a tissue of central
importance because it contains approximately three-
fourths of the body’s intracellular water volume (ICV)
and one-third the total interstitial fluid volume (IFV).
How much water is absorbed from skeletal muscle when
plasma osmolality increases? Previous investigations in
isolated muscle (15, 19, 27) have not studied steady-
state changes but instead have focused on the first sev-
eral seconds of the osmotic transient. Such studies have
attempted to calculate the initial transcapillary osmotic
pressure gradient after osmolality is increased, then
compare that gradient to what would be expected for an
ideal membrane obeying van’t Hoff’s law. However,
there is some disagreement on how much of an osmotic
*effect small molecules (the size of electrolytes and simple
sugars) have across the capillary wall (17, 27). The un-
certainties of the water flux between tissue and plasma
caused by small molecules, the lack of previous measure-
ments of ICV and IFV changes, and the possible changes
in hydrostatic and colloid osmotic pressures have made
R1318 0363-6119/93 $2.00 Copyright
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AND PHILIP D. WATSON
University of South Carolina,
models of the absorption of water from tissue very com-
plex (2,5,7,14,16,25,26). There is no consensus among
these models as to whether the water redistribution is
caused directly by a transcapillary osmotic gradient or
indirectly by other pressures.
The purpose of the current investigation was to de-
scribe quantitatively the relationship between plasma
osmolality and total tissue water (TTW) in skeletal
muscle under steady-state conditions. To describe the
fluid shift, we have adapted a useful approach that has
previously been used to describe the behavior of a vari-
ety of cell types placed in anisosmotic media (13). The
approach is to compare the observed steady-state vol-
ume changes to those predicted by ideal behavior when
all of the water volume is “osmotically active.,, An im-
portant component of our study was to determine the
relationship between osmolality and IFV.
METHODS
Perfusion preparation. Experiments were performed in a calf
musclepreparation (22) isolatedfrom 15 fasted cats (6 males,9
females)weighing between 2 and 4 kg. Animals were anesthe-
tized with 38 mg/kg pentobarbital sodiumbefore surgery. The
thigh musclesand skin covering the calf wereremovedwith heat
cautery, without cutting, burning, or ligating any of the calf
muscles.The popliteal artery and vein were cannulated closeto
the knee, and flow to the foot was tied off at the ankle. The
preparation was weighed before perfusion and then wrapped
with plastic food wrap to minimize evaporation. Blood was
taken from the samecat and diluted in well-dialyzed bovine
serumalbumin and electrolytes (21) to give an initial pefisate
volume of 40-60 ml with an averagehematocrit of 21 t 4%. The
recirculating perfusate waspumped (Harvard model 1215peri-
staltic pump) at a constant flow rate through a filter and bubble
trap and kept warm at 37-38OC. The reservoir was mixed and
oxygenated by bubbling with saturated 95% 02-5% CO,. Arte-
rial and venouspressureswere measuredthrough branchedtub-
ing adjacent to the cannulasusing Statham pressuretransduc-
ers. Profound vasodilation induced by papaverine (10s5M final
extracellular concn) and isoproterenol (10m7M) prevented pos-
sible changesin vascular tone and vascular pressuresand main-
tained normal vascular permeability (23). During the first sev-
eral minutes of perfusion 2-4 ml of fluid was filtered by
increasing flow and raising the height of the venous cannula.
Then venouspressurewas set by adjusting the height of venous
outflow until the muscle was isogravimetric. The baseline
weight increasecausedby leaksinto the plastic wrap and on the
scale is smaller in this preparation than in the more familiar
hindlimb model. Nevertheless,the changesin weight causedby
osmolality wereadjustedfor the baselineweight change(average
0.02 g/min).
Osmotic transient procedure. Osmolality was increasedby
adding smallvolumes(0.4-2.0 ml) of 2.8 M saline(26 transients
in 10 preparations) or 2.2 M sucrose(8 transients in 5 prepa-
rations) into the reservoir. An osmolality decrease(water addi-
tion) precededNaCl addition in one preparation. Transcapillary
0 1993 the American Physiological Society
 FLUID REDISTRIBUTION DURING HYPEROSMOLALITY R1319
fluid shifts (changesin TTW) were recorded continuously by osmolality was increasedwere compared with repeated-meas-
placing the muscleon a sensitive balancemodified with a force ures analysis of variance. Data are presentedas means+ SE.
transducer (Statham UC-2) connectedto a Gould chart recorder
with a sensitivity of 0.5 g/cm pen deflection. After decreasesin RESULTS
weight were complete, osmolality was determined again. Muscle weight averaged 53.0 t 4.9 g and was perfused
Tracer-measured fluid redistribution. IFV was calculated in
eight preparations by subtraction of Evans blue dye (EBD) with a blood flow of 37 t 4 ml min-l 100 g-l. Osmolality
l l

plasmavolume from extracellular volume measuredwith either
51Cr-labeledEDTA (n = 5) or [3H]mannitol (n = 3). Tracers
weresampledfrom the reservoir of the recirculating perfusate.A
plasma blank and standards were made from samplestaken
after the initial filtration procedures described above. Addi-
tional l-ml sampleswere taken 90 min after tracers were added
and again - 17, 40, and 65 min after hypertonic solutions were
added. Corrections were made for tracer removed during sam-
pling.
EBD stock wasprebound to albumin and filtered before use,
and 500 ~1was addedto the reservoir 15 min after the start of
perfusion. This volume gave an absorbanceof 0.5-1.0 U at 620
nm in plasmasamplesdiluted 26-fold in cyanomethemoglobin
solution. The 620-nm readingwascorrected for possibleplasma
hemoglobinby comparisonof the 420- and 620-nm absorbance
ratios of sampleand standards. The correction for hemolysis
causedby blood pumping was never greater than 5%.
Enough 51Cr-EDTA was added with EBD to give at least
20,000 counts/l00 ~1 plasmain an automated gammacounter.
To account for radioactive decay of 51Cr (half-life, 28.5 days),
each set of sampleswas counted with aged standardsfrom the
samestock of tracer. In mannitol studies,0.5-1.0 ml unlabeled
6% mannitol was added to the perfusate to increasemannitol
concentration to -5 mM. Radioactivity in IOO-~1[3H]mannitol
plasma samples,counted for 20 min in an automated scintil-
lation counter, was sufficient to give at least 100,000
disintegrations min-l (dpm) ml-l. [3H]mannitol was cor-
l l
was 324 t 4 mosmol/kgH20 before the osmotic tran-
sients, which approximates in vivo cat plasma measured
by others (9).
Vascular-pressure stability and weight change. As
shown in Fig. 1, arterial and venous pressure changes
were negligible, ranging between 0 and 2 mmHg in venous
pressure and between 0 and 10 mmHg in arterial pressure
for all osmolality increases studied. In the example, the
decrease in weight was complete by 7 min and averaged
12 min.
TTPV. Figure 2 is a plot of changes in TTW measured
from weight as a function of the increase in plasma os-
molality. Osmolality was increased with step sizes rang-
ing between 17 and 129 mosmol/kgH20. The largest in-
creases in osmolality (-twofold increase after several
large NaCl transients) caused ~32 ml/100 g decreases in
TTW (Fig. 2).
The fit of the TTW data to the ideal osmometer model
is graphically illustrated in Fig. 2, inset, where AV is
plotted against 01/02 - 1. Equation 2 predicts that when
AV is plotted against 01/02 - 1, the data should pass
through the origin and lie on a straight line with slope of
V1. As seen, the slope is linear (r = 0.995), and least-
squares fit to the data gave a V1 of 65 t 1 ml/l00 g with
95% confidence intervals of 63-68 ml/100 g. The linear
slope is consistent with a tissue osmotic mass (2M) inde-

rected for quenching with appropriate standards. Change in

ICV wascalculated from differences in TTW and IFV. ICV was pendent of plasma osmolality, averaging 20 t 1
calculated before osmotic transients by subtracting IFV and mosmol kgHZO-ll 100 g-l.
l

tissue dry weight from total tissue weight (n = 4). Dry weight In addition to the 34 increasing osmotic steps, in one
was determinedby drying a largeportion of the musclemassto preparation distilled water was added to the reservoir of a
a constant weight at 60°C.
Plasmawas separatedby centrifugation, and plasmaosmo-
lality was determined by freezing-point depression(Osmette, t
Precision Systems). Plasma sodium was determined by flame 25 mmHg
photometer in samplesfrom five experiments. -fji!
-- 7 A-
Perfect osmometer model. The decreasesin TTW were de-
scribed with the following massbalance equation. If muscle
osmolar mass(M) remains constant during an osmotic tran-
sient then
O,V, = O,v, = M (0 t
10 mmHg
where 0 is osmolality and V is tissuevolume participating in the J
fluid shifts during steady-stateisotonic ( 01, V,) and hypertonic
(0,, V,) conditions. Assuming the density of water is 1 g/ml, 1 minute
l-l
then the weight lossduring an osmotictransient, AV, is equalto
v2 - V1. Thus, if muscletissue behaveslike a perfect osmom- T
eter, the following equation should adequatelydescribesteady- 1 gram
state weight changesfor any changein osmolality: 1

AV=V,@l) 01
2
Statistics.Results are presentedper 100 g of musclebefore
perfusion. Muscle weight was determined by subtracting the
weight of nonmuscletissue dissectedat the end of perfusion
(primarily the tied-off foot and bones) from the weight of the
preparation at the start of an experiment. Significance wasset
at P < 0.05. BaselineIFV measurementswith the two extracel-
lular tracers were comparedwith a t test. Changesin IFV after
(2)
0.5 ml hypertonic saline
Fig. 1. Chart recording of weight and blood pressure changes in an
example muscle weighing 52.4 g. Osmolality was increased by adding a
bolus of 2.2 M NaCl into a well-mixed reservoir. Steady-state plasma
osmolality was increased by 39 mosmol/kgH,O with hypertonic saline.
Decreased muscle weight reflects transcapillary absorption as total tis-
sue water (TTW) decreases. Mean arterial and venous pressures at the
cannulas were 45 and 4 mmHg, respectively.

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 R1320 FLUID REDISTRIBUTION DURING HYPEROSMOLALITY

0 20 40 60 80 100
I- ’ I I I I -I
- CD -5-
80
t!P 400
g
-10 -
@.
K 4) II Plasma osmolality
II I
0
E
w-15 - 0 cn ,
II
3 3 f 350
II I
I- 00
I- -20 -
0 II
o-c 0 II I
a> -25 - II
0 -&
300 I
1 I I I I I I

G l 0
I I 1 I
-35
1 .o 1.5 2.0

Osmolality ratio (02/0,)

Fig. 2. TTW and plasma osmolality changes (0,/O,) during 34 osmotic
transients from an average starting osmolality of 324 t 4 mosmol/
kgH,O. 0, NaCl; 0, sucrose. Inset, change in TTW vs. 0,/O, - 1. Re-
gression line described in text. 2 -10

red blood cell-free perfusate to lower osmolality from 324

to 255 mosmol/kgH20. Lowering osmolality increased
TTW 18.9 ml/100 g, which corresponded to a V1 of 69.7 0 20 40 60 80 100
ml/100 g. The osmolality was then returned from 255 to Time (minutes)
325, with saline causing a 14.97 ml/l00 g decrease in Fig. 3. TTW, interstitial fluid volume (IFV), and plasma osmolality
TTW, which was consistent with a V1 of 69.5 ml/100 g. changes in an example preparation that had the greatest change in IFV.
IFV and ICV. IFV was 18.4 t 2.3 ml/l00 g before the Change in IFV was small in comparison to initial IFV (23.8 ml/100 g).
osmotic step. EDTA (n = 5) and mannitol (n = 3) gave Osmolality and TTW were stable after 17 min.

indistinguishable initial IFV values (19.0 t 3.3 ml/100 g

and 17.4 t 4.1 ml/l00 g, respectively), and IFV was 5 I I I I I
changed very little by osmolality during any of the prepa-
rations. 0
0
Figure 3 shows results for the preparation with tracers
that had the largest change in IFV. Although osmolality 0 9
0
(and ICV) changes were greater in this preparation than t
in the others, IFV changes were small, ranging between v 0
-3.7 and 0.7 ml/l00 g (-16% to 3% change from an
initial IFV of 23.8 ml/100 g). VP
v v
Figure 4 shows the relationship between IFV and ICV
V
17 min after osmolality was changed. Although ICV 0; -10
~
clearly was reduced with increases in osmolality, IFV was z
independent of osmolality. Figure 5 shows the stability bz
0 IFV (NaCI) v
after 17 min. There was no statistical change in IFV at
o-

V ICV (NaCI) V
-15
any of the time points for either NaCl and sucrose or Fi 0 IFV (Sucrose)
CI
when combined as one group. The average change in IFV u v ICV (Sucrose)
-c
for both solutes was -0.1 t 0.7 ml/100 g at 17 min (n = 0
lo), 0.8 t 0.5 at 40 min (n = 8), and 0.9 t 1.0 at 65 min -20
0 20 40 60 80 100 Go
(n = 5). However, ICV decreased during the first 17 min
Change in Osmolality (mOsm/kg)
and remained stable thereafter. Consistent with the
stable ICV after 17 min, plasma sodium concentration Fig. 4. Changes in plasma osmolality, IFV, and intracellular water vol-
ume (ICV) 17 min after solutes were added to plasma.
only changed -1.2 t 1.1 (n = 5) meq/l between 17 and 40
min.
In four preparations from the dry tissue data we calcu- DISCUSSION
lated ICV at the end of the baseline period to be 59,61,63, We can conclude without the use of a model that
and 68 ml/l00 g (mean 62.7). These values are reduced 2 steady-state IFV did not change when ICV and TTW
ml/l00 g if the small blood volume in muscle is subtracted decreased after a wide range of osmolality increases
(within and bevond the nathonhvsiological range). To our

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 FLUID REDISTRIBUTION DURING HYPEROSMOLALITY R1321

I I I I I osmolality. Because of the requirement for extracellular

l IFV (NaCI)
0 IFV (Sucrose)
V This suggests that little cell water is bound in cat skeletal
v ICV (NaCI)
I I I I
10 20 30 40 50
Time (minutes)
Fig. 5. Changes in IFV (broken lines) and ICV (solid lines) 17, 40, and
65 min after osmolality was increased. ICV, but not IFV, decreased
significantly during the first 17 min. Neither volume changed signifi-
cantly after 17 min.
knowledge, there are no previous osmotic weight tran-
sient studies in skeletal or cardiac muscle measuring IFV
and ICV. Additionally, there are no previous studies re-
porting the relationship between TTW and graded
plasma osmolality changes in isolated tissues, but Arieff
and Kleeman (1) did measure skeletal muscle volume
shifts in intact rabbits subjected to glucose-induced hy-
perosmolality. When their data are converted to
milliliters/100 g of initial weight+ they find that an os-
molality change of 14.8% caused a muscle TTW change of
8.4 ml/100 g, compared with 8.7 calculated from the data
in Fig. 2. In that same study, hyperglycemia did not
change tissue sodium or chloride content, consistent with
our finding of no change in IFV. Thus, despite the pos-
sibility for multiple factors to influence skeletal muscle
fluid balance in vivo, the results of Arieff and Kleeman
are similar to those in the current study.
This and previous osmotic weight studies (19,27) have
been able to measure rapid changes in TTW gravimetri-
tally with good precision (Fig. 1) and low variability be-
tween different preparations (Fig. 2). However, descrip-
tion of the interaction between IFV and ICV is required
for complete understanding of fluid balance when osmo-
lality changes. Little is known about this interaction,
perhaps because of the technical difficulty in measuring
fluid shifts with tracers. One problem is a relatively low
sensitivity for the measurement of IFV changes in com-
parison to TTW. We attempted to avoid this problem by
measuring IFV of the entire muscle mass rather than the
usual sampling of small pieces of tissue as required in vivo
(1, 24). We also attempted to amplify possible changes in
IFV by inducing several large changes in osmolality and
ICV. Figure 4 clearly shows that IFV was not changed by
1 Arieff and Kleeman reported % water in tissue biopsies (mg/lOO g).
By mass balance, the change in TTW per 100 g of initial weight can be
obtained from ATTW = 100(R2 - R,)/(lOO - R,), where R, and Rz are
the initial and % water in biopsy.
tracer equilibration, changes in IFV and ICV could not be
measured until the fluid shift was complete. However,
IFV was stable when measured, and changes in TTW
were monotonic. Thus if there was a rapid and transient
change in IFV during the fluid shift it was probably small
in comparison to the decrease in TTW and ICV.
V1, the amount of TTW that apparently participated in
fluid shifts, was 65 t 1 ml/l00 g. This was very close to
ICV measured in this study, 62.7 ml/l00 g, and to the 65
ml/l00 g calculated from the data of Wiig and Reed (24).
muscle fibers and that if volume regulatory increase
mechanisms are important they are either very slow act-
ing or offset by unidentified processes. Similar results
have been found in frog muscle when similar distribution
I I
volume methods were used (6). However, studies using
microscopic measurements in frog muscle conclude ICV
60 70
decreases less than expected (3, 4, 6). This inconsistency
was explained by concluding that the inactive volume was
limited to the sarcoplasmic reticulum, which swelled
rather than decreased like the rest of the fiber. Even if the
sarcoplasmic reticulum behaves oddly, it is probably less
important to fluid shifts in mammalian skeletal muscle
because this space occupies only - 1% of ICV in cats (18)
vs. -13% in frogs (3).
What pressures were involved in decreasing TTW?
Other than small-molecule osmotic pressure, the other
pressures that move water across the capillary wall are
the hydrostatic and colloid osmotic pressures described
by the Starling-Landis equation (11). We set the condi-
tions of the current investigation to avoid significant con-
tributions from the Starling pressures. Initial IFV was set
above normal at 17-18 ml/100 g by filtration (see RE-
SULTS). When IFV is greater than 12 ml/l00 g, intersti-
tial hydrostatic pressure (Pif) does not change with fur-
ther increases in IFV (24). Note that TTW is decreasing
in this study, so that tissue pressure is most unlikely to
increase. In addition to effects on Pif and compliance, a
larger-than-normal IFV dilutes interstitial protein and
reduces possible interstitial colloid osmotic pressure
(COPif) changes. Hence, our initial tissue hydration made
the possible magnitude of changes in Pif and COPif
caused by subsequent IFV changes very small. Further-
more, because there was no evidence that IFV changed, it
is very unlikely that Pif or COPif contributed to fluid
shifts in this study. Capillary hydrostatic pressure (P,,,)
could not have changed significantly, because vascular
pressures and blood flow were held constant by the ex-
perimental conditions. Finally, there was an unavoidable
dilution of plasma protein during the water shift from
tissue to plasma. This would tend to increase IFV and
offset transcapillary absorption by reducing plasma COP.
However, calculations using the Landis and Pappen-
heimer equation for osmotic pressure (11) showed plasma
COP decreased <5 mmHg even during the largest tran-
sient in Figs. 4 and 5. The slight trend for IFV to increase
after 17 min (see RESULTS), although insignificant, was
consistent with a slight decrease in plasma COP. In sum-
mary, it is unlikely that the fluid shifts reported in this
paper were significantly affected by any of the Starling

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R1322 FLUID REDISTRIBUTION
pressures and, therefore, only small-molecule osmotic
pressures can explain the decreases in TTW.
How does plasma hyperosmolality decrease ICV with-
out changes in IFV? Because the osmotic reflection co-
efficient (a,,,) for small solutes at the capillary wall is 0.5
or greater (27), the increase in mosmolality (17-129
mosmol/kgHZO) produces a large and rapid increase in
transcapillary osmotic pressure of 160- 1,240 mmHg. This
starts a rapid water flow across the capillary wall. Because
small solutes are producing the osmotic pressure, this
pressure must act at transcapillary pathways that largely
exclude small solute so that the transcapillary flow leaves
the interstitial solute behind. This concentration polar-
ization or sieving is sometimes called “solute buffering”
(8). Consequently, interstitial osmolality rises rapidly
even without the diffusion of solute from plasma. The
increase in interstitial osmolality then pulls water from
cells, preventing a large decrease in IFV and buffering the
interstitial changes. Diffusion of the osmotic solute from
plasma adds to the interstitial osmolality, slowing the
transcapillary water flow and increasing the transsarco-
lemma1 flow.
The steady-state IFV is determined by ccap and other
permeability parameters. If CT,,~were 1.0, then the entire
tissue would behave like a perfect osmometer, with both
IFV and ICV shrinking in proportion to the osmotic step.
If CT,,~were zero, then there would be little or no trans-
capillary flow and IFV would increase by the amount of
the ICV decrease. Hence, as IFV was unchanged, gcap
must lie somewhere in the middle. A precise estimate of
the value of ccap giving an unchanged IFV will take a
complicated analysis involving water and solute perme-
ability of the capillary wall, blood flow rate, and the water
permeability of the muscle cells, which is beyond the
scope of this paper. The fact that changes in IFV can be
in either direction, depending on CT,,~, may explain why
studies modeling osmotic transients with NaCl or sucrose
in the isolated perfused heart (2,5) or whole body (14,26)
disagree even in the direction of IFV changes.
A previous study in the isolated lung (20) also found
that IFV did not change during osmotic transients. Al-
though this finding is similar to ours, the interpretation
of the process in the lung was in sharp contrast to what
we feel explains our findings. The authors of the previous
study explicitly assumed that ccap for small molecules was
essentially zero. They argued that NaCl rapidly equili-
brated into the interstitial space, thereby causing water
flow from cells to increase Pif enough to rapidly force the
extra fluid into the capillary lumen. Because cell volume
and TTW must have decreased, it is hard to see how a
large increase in Pif could occur. The belief that hyper-
osmolality indirectly decreases TTW because of increased
Pif or decreased interstitial COP is shared by others mod-
eling fluid balance in both the whole animal (7, 14, 16)
and in the isolated perfused heart (5). Although it is
beyond the scope of this paper to delineate differences
between skeletal muscle and other tissues, it important to
emphasize that, other than the cited lung study (20),
previous studies modeled changes in IFV rather than di-
rectly measuring this space.
The finding of no change in IFV, together with the
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DURING HYPEROSMOLALITY
increase in interstitial osmolality, means that the inter-
stitial osmolar mass was increased by the osmotic bolus.
However, equation 2 was obtained assuming that there
was no tissue osmolar mass change and yet AV was still a
linear function of the osmolality change as predicted (Fig.
2, inset). This inconsistency is explained by the following.
Let the tissue osmolar mass change be AM, where VZOg -
w = AM and Vi is the initial tissue fluid volume. Re-
arranging and defining AV = V2 - Vi, we obtain
0
AV=Vi(~- l)- (3)
2
Because IFV is constant, AM = IFV(OZ - 0,); therefore
AV=Vi($ 0 - 1) - IFV ($0 - 1) (4)
2 2
which reduces to
0
AV = (Vi - IFV)( $ - 1) (5)
2
Equation 3 shows that a linear function is obtained when
part of the tissue volume (IFV) does not change but does
reach diffusion equilibrium with the osmoles. Equation 2
is a special case of equation 3.
AS discussed above, (Vi - IFV) was found to be 65
ml/100 g, which was not different from cell volume. Be-
cause IFV was 18, this is consistent with Vi representing
the initial TTW of 76 ml/l00 g hydrated by 7 ml/l00 g, as
described in METHODS. Hence, the lack of change in IFV
is consistent with both the linear relationship between
AV and 01/02 - 1 and the measured volume of ICV.
In conclusion, after rapid osmotic transient with hy-
pertonic NaCl and sucrose, steady-state IFV was un-
changed and the decreases in TTW and ICV were similar.
These findings imply that when steady state is reached,
the total transcapillary water flow is equal to the total
transsarcolemmal flow, so that plasma volume expansion
is approximately equal to cell volume shrinkage.
Perspectives
It is well established that blood volume influences car-
diovascular performance and that sometimes both fluid
homeostasis and cardiovascular function are stressed si-
multaneously. Heat exposure, exercise, and hemorrhagic
shock are normal and pathological examples. Interest-
ingly, in all of these conditions there are significant in-
creases in plasma osmolality that are graded with the
severity of the condition. The present findings are of
great relevance here. Using equation 2 and the value V1 =
65 ml/l00 g, we can show that if extracellular solutes have
increased plasma osmolality from 300 to 330 mosmol/
kgH20, then skeletal muscle must have contributed 59
ml/kg (l-2 liters of fluid in the average person) toward
maintaining the plasma volume. Hence, fluid shifts
driven by small-molecule osmotic pressure could be as
important as the Starling pressures in the maintenance of
cardiovascular performance under some circumstances.
The present findings in isolated muscle should be com-
pared with changes in the skeletal muscle of intact ani-
mals to determine the relative contributions of the Star-
ling pressures and small-molecule osmotic pressure to the
 FLUID REDISTRIBUTION DURING HYPEROSMOLALITY R1323

maintenance of plasma volume under a variety of condi- 11. Landis, E. M., and J. R. Pappenheimer. Exchange of sub-
tions. For instance, plasma COP may increase and Pcap stances through capillary walls. In: Handbook of Physiology. Cir-
culation. Bethesda, MD: Am. Physiol. Sot., 1963, sect. 2, vol. 2,
may decrease during dehydration because of the reduc- chapt. 24, p. 961-1034.
tion in plasma volume. Both changes tend to enhance 12 Lundvall, J., S. Mellander, H. Westling, and H. White.
fluid absorption and decrease IFV. However, plasma COP l
Fluid transfer between blood and tissues during exercise. Acta
and
move in the other direction and tend to reduce
p,,
Physiol. Stand. 85: 258-269, 1972.
fluid absorption during hypertonic saline resuscitation. A 13. MacKnight, A. D. C., and A. Leaf. Regulation of cell volume.
useful starting point would be to compare the water shifts Physiol. Rev. 57: 510-573, 1977.
measured by biopsy samples taken from intact animals to 14. Mazzoni, M. C., P. Borgstrom, K. E. Arfors, and M. Intag-
the changes predicted by the results presented here. lietta. Dynamic fluid redistribution in hyperosmotic resuscita-
tion of hypervolemic hemorrhage. Am. J. Physiol. 255 (Heart Circ.
Physiol. 24): H629-H637, 1988.
We wish to thank Evelyn B. May for her excellent technical assis-
tance. 15. Pappenheimer, J. R., E. M. Renkin, and L. M. Borrero.
This study was supported by National Heart, Lung, and Blood In- Filtration, diffusion, and molecular sieving through the peripheral
stitute Grant HL-24314 and by the South Carolina Affiliate of the capillary membranes. A contribution to the pore theory of capil-
American Heart Association. lary permeability. Am. J. Physiol. 167: 13-46, 1951.
Present address of M. T. Hamilton: Dept. of Physiology and Cell 16. Pirkle, J. C., and D. S. Gann. Restitution of blood volume after
Biology, Univ. of Texas Medical School, Houston, TX, 77225. hemorrhage: mathematical description. Am. J. Physiol. 228: 821-
Address for reprint requests: P. D. Watson, Dept. of Physiology, 827, 1975.
Univ. of South Carolina, Columbia, SC 29208. 17. Rippe, B., and B. Haraldsson. Capillary permeability in rat
Received 11 February 1993; accepted in final form 30 April 1993. hindquarters as determined by estimations of capillary reflection
coefficients. Acta Physiol. Stand. 127: 289-303, 1986.
REFERENCES 1Q
IV. Schmalbruch, H. The membrane systems in different fibre types
1. Arieff, A. I., and C. R. Kleeman. Studies on mechanisms of of the triceps surae muscle of cat. CeZZTiss. Res. 204: 187-200,
cerebral edema in diabetic comas. J. Clin. Invest. 52: 571-583, 1979.
1973. 19. Vargas, F. F., and J. A. Johnson. An estimate of reflection
2. Bassingthwaighte, C. A. Goresky.
J. B., and Modeling in the coefficients for rabbit heart capillaries. J. Gen. Physiol. 47: 667-
analysis of solute and water exchange in the microvasculature. In: 677, 1964.
Handbook of Physiology. The Cardiovascular System. Microcircu- 2. Wangensteen,D., H. Bachofen, and E. R. Weibel. Lung
&ion. Bethesda: Am. Physiol. Sot., 1984, sect. 2, pt. 1, vol. 4, ’
tissue volume changes induced by hypertonic NaCl: morphometric
chapt. 13, p. 549-626. evaluation. J. Appl. Physiol. 51: 1443-1450, 1981.
3. Birks, R. I., and D. F. Davey. Osmotic response demonstrating
the extracellular character of the sarcoplasmic reticulum. J. 21* Watson, P. D. Colloid osmotic pressure changes in isolated iso-
Physiol. Lond. 202: 171-188, 1969. gravimetric cat hindlimb. Am. J. Physiol. 242 (Heart Circ. Physiol.
11): H512-H519, 1982.
4. Blinks, J. R. Influence of
osmotic strength on cross-section and
volume of isolated single muscle fibres. J. Physiol. Lond. 177: 22. Watson, P. D., R. P. Garner, and D. S. Ward. Water uptake
42-57, 1965. in stimulated cat skeletal muscle. Am. J. Physiol. 264 (Regulatory
5. Bloom, A model for osmotically induced
G., and J. A. Johnson. Integrative Comp. Physiol. 33): R790-R796, 1993.
weight transient in the isolated rabbit heart. Microvasc. Res. 22: 23. Watson, P. D., M. B. Wolf, and I. S. Beck-Montgomery.
64-79, 1981. Blood and isoproterenol reduce capillary permeability in cat hind-
6. Dydynska, M., and The osmotic properties of
D. R. Wilkie. limb. Am. J. Physiol. 252 (Heart Circ. Physiol. 21): H47-H53,
striated muscle fibres in hypertonic solutions. J. Physiol. Lond. 1987.
169: 312-329, 1963. 24. Wiig, H., and R. K. Reed. Interstitial compliance and transcap-
7. Gann, D. S., D. E. Carlson, G. I. Byrnes, J. C. Pirkle, and illary Starling pressures in cat skin and skeletal muscle. Am. J.
C. F. Allen-Rowlands. Role of solute in the early restitution of Physiol. 248 (Heart Circ. Physiol. 17): H666-H673, 1985.
blood volume after hemorrhage. Surgery 94: 439-446, 1983.
Wolf, M. B. Estimation of parameters affecting rapid fluid trans-
8. Grabowski, E. F., and J. B. Bassingthwaighte. An osmotic 25* fers in the whole body. I. Isotonic infusions. Ann. Biomed. Eng. 3:
weight transient model for estimation of capillary transport pa- 209-224, 1975.
rameters in myocardium. In: Microcirculation, edited by J. Gray-
son and W. Zingg. New York: Plenum, 1976, vol. 2, p. 19-50. 26. Wolf, M. B. Estimation of whole-body capillary transport param-
9. Jarhult, J., J. Holmberg, J. Lundvall, and S. Mellander. eters from osmotic transient data. Am. J. Physiol. 242 (ReguZatory
Hyperglycemic and hyperosmolar responses to graded hemor- Integrative Comp. Physiol. 11): R227-R236, 1982.
rhage. Acta Physiol. Stand. 97: 470-475, 1976. 27. Wolf, M. B., and P. D. Watson. Measurement of the osmotic
10. Klein, J. R. Estimation of blood in tissue. Arch. Biochem. Bio- reflection coefficient for small molecules in the cat hindlimb. Am.
phys. 8: 421-424, 1945. J. Physiol. 256 (Heart Circ. Physiol. 25): H282-H290, 1989.

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