86:1092-1096, 1999.

; J Appl Physiol
Ethan R. Nadel
Nina S. Stachenfeld, Loretta DiPietro, Cheryl A. Kokoszka, Celso Silva, David L. Keefe and
young women
Physiological variability of fluid-regulation hormones in
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rap id co m m u n icatio n
Physi ol ogi cal vari abi l i ty of ui d-regul ati on
hormones i n young women
NI NA S. STACHENFELD,
1
LORETTA DI PI ETRO,
1
CHERYL A. KOKOSZKA,
1
CELSO SI LVA,
4
DAVI D L. KEEFE,
4
AND ETHAN R. NADEL
1,2,3
1
The J ohn B. Pierce Laboratory, Departments of
2
Epidemiology and Public Health,
and
3
Cellular and Molecular Physiology, Yale University School of Medicine, New Haven,
Connecticut 06519; and
4
Women and I nfants Hospital, Brown University School of Medicine,
Providence, Rhode I sland 02905
Stachenfeld, Nina S., Loretta DiPietro, Cheryl A.
Kokoszka, Celso Silva, David L. Keefe, and Ethan R.
Nadel. Physi ol ogi cal vari abi l i ty of ui d-regul ati on hormones
i n young women. J . Appl. Physiol. 86(3): 10921096,
1999.We tested the physi ol ogi cal rel i abi l i ty of pl asma reni n
acti vi ty (PRA)and pl asma concentrati ons of argi ni ne vasopres-
si n (P
[AVP]
), al dosterone (P
[ALD]
), and atri al natri ureti c pepti de
(P
[ANP]
) i n the earl y fol l i cul ar phase and mi dl uteal phases
over the course of two menstrual cycl es (n 9 women, ages
25 1 yr). The rel i abi l i ty (Cronbachs 0.80) of these
hormones wi thi n a gi ven phase of the cycl e was tested 1) at
rest, 2) after 2.5 h of dehydrati ng exerci se, and 3) duri ng a
rehydrati on peri od. The mean hormone concentrati ons were
si mi l ar wi thi n both the earl y fol l i cul ar and mi dl uteal phase
tests; and the mean concentrati ons of P
[ALD]
and PRA for the
three test condi ti ons were si gni cantl y greater duri ng the
mi dl uteal compared wi th the earl y fol l i cul ar phase. Al though
Cronbachs for resti ng and recovery P
[ANP]
were hi gh (0.80
and 0.87, respecti vel y), the resti ng and rehydrati on val ues for
P
[AVP]
, P
[ALD]
, and PRA were vari abl e between tri al s for the
fol l i cul ar ( from 0.49 to 0.55) and the l uteal phase ( from
0.25 to 0.66). Physi ol ogi cal rel i abi l i ty was better after dehy-
drati on for P
[AVP]
and PRA but remai ned l ow for P
[ALD]
.
Al though resti ng and recovery P
[AVP]
, P
[ALD]
, and PRA were
not consi stent wi thi n a gi ven menstrual phase, the di ffer-
ences i n the concentrati ons of these hormones between the
di fferent menstrual phases far exceeded the vari abi l i ty wi thi n
the phases, i ndi cati ng that the l ow wi thi n-phase rel i abi l i ty
does not prevent the detecti on of menstrual phase-rel ated
di fferences i n these hormonal vari abl es.
al dosterone; reni n; atri al natri ureti c pepti de; argi ni ne vaso-
pressi n; estrogen; progesterone
THE BODYS WATER- AND SODI UM-regul ati ng hormones
vary consi derabl y over the course of the menstrual
cycl e (79, 14, 15, 18, 19). For exampl e, duri ng the
mi dl uteal phase of the menstrual cycl e, pl asma al doste-
rone concentrati on (P
[ALD]
) and pl asma reni n acti vi ty
(PRA) are greater at rest (9) and duri ng exerci se (14,
15) than i n the fol l i cul ar phase. I n addi ti on, resti ng
pl asma argi ni ne vasopressi n concentrati on (P
[AVP]
) i s
hi gher (8)duri ng the preovul atory and mi dl uteal phases
of the cycl e when pl asma estrogen concentrati on (P
[E
2
]
)
i s hi gh. I n l ower ani mal s, estrogen admi ni strati on
i ncreases osmoti c sti mul ati on of AVP (1, 4, 5) and water
retenti on (3), and both estrogen and progesterone ex-
hi bi t i mportant effects on sodi um regul ati on and the
sodi um-regul ati on hormones (1012, 21); thi s supports
the hypothesis that the gonadal steroids have important
modulatory effects on body uid and electrolyte balance.
No studi es exi st that exami ne the physi ol ogi cal rel i -
abi l i ty of the ui d-regul ati ng hormones wi thi n a gi ven
phase and over the course of two or more menstrual
cycl es. Reported pl asma concentrati ons of these hor-
mones across di fferent menstrual cycl es di ffer due to
natural physi ol ogi cal vari ati ons, due to sel ecti on of an
i nappropri ate day to conduct physi ol ogi cal testi ng, due
to vari ati ons i n water and/or sodi um i ntake, or due to
i naccurate hormone-anal ysi s techni ques. The purpose
of thi s study was to el i mi nate vari abi l i ty caused by the
l atter three reasons to determi ne the natural physi ol ogi -
cal vari abi l i ty of the responses of ui d- and sodi um-
regul ati ng hormones over two menstrual cycl es. Accord-
i ngl y, we tested women twi ce duri ng the earl y fol l i cul ar
phase (when estrogen and progesterone are l ow) and
twi ce duri ng the mi dl uteal phase of the menstrual cycl e
(when estrogen and progesterone are hi gh).
METHODS
StudyDesign
Subjects were ni ne heal thy, nonsmoki ng women (age, 25
1 yr; range, 2231 yr). To dri ve the ui d-regul ati on system,
each woman parti ci pated i n a seri es of dehydrati on experi -
ments i n whi ch the study hormones were measured 1) at rest,
2) duri ng dehydrati on, and 3) duri ng rehydrati on i n both the
earl y fol l i cul ar and the mi dl uteal menstrual phases. The
study desi gn empl oyed four dehydrati on experi ments: two
conducted i n the earl y fol l i cul ar phase (24 days after the
begi nni ng of menstrual bl eedi ng) and two i n the mi dl uteal
phase of the menstrual cycl e (conducted 710 days after the
l utei ni zi ng hormone peak), as determi ned i ndi vi dual l y by the
use of ovul ati on-predi cti on ki ts (OvuQui ck; Qui del , San Di ego,
CA). The tests were conducted duri ng nonconsecuti ve men-
The costs of publ i cati on of thi s arti cl e were defrayed i n part by the
payment of page charges. The arti cl e must therefore be hereby
marked advertisement i n accordance wi th 18 U.S.C. Secti on 1734
sol el y to i ndi cate thi s fact.
1092 http://www.jap.org

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strual phases, 1216 wk apart. To veri fy the phase of the
menstrual cycl e, pl asma l evel s of 17-estradi ol and progester-
one were assessed from a basal bl ood sampl e.
Dehydration Experiments
On the day of the dehydrati on test, vol unteers arri ved at
the l aboratory between 7:00 and 8:00 AM, after they had
eaten onl y a prescri bed l ow-fat breakfast (300 kcal ). The
subjects refrai ned from al cohol and caffei ne for 12 h before
the experi ment. Subjects were asked to dri nk 7 ml /kg body
wei ght of tap water at home before arri val at the l aboratory.
On arri val at the l aboratory, the subjects gave a basel i ne
uri ne sampl e; they were wei ghed and then sat on the contour
chai r of a cycl e ergometer i n the test chamber [27C, 30%
rel ati ve humi di ty (RH)] for 60 mi n of control rest. Duri ng the
control peri od, an i ndwel l i ng catheter was pl aced i n an arm
vei n. At the end of the control peri od, a 20-ml bl ood sampl e
was drawn and uri ne was col l ected. Consi stency of the pretest
hydrati on state was assessed from the speci c gravi ty of the
basal uri ne sampl e (mean 1.001), whi ch di d not di ffer
across tri al s.
After the control peri od, the chamber temperature was
i ncreased to 36C. The subjects then exerci sed at 50% maxi -
mal power output for 150 mi n, wi th 5-mi n rest peri ods every
25 mi n, duri ng whi ch ti me they were depri ved of ui ds. Bl ood
sampl es (1020 ml ) were drawn and body wei ght was as-
sessed at 60, 120, and 150 mi n duri ng exerci se. At the end of
exerci se, the chamber temperature was reduced to 27C.
After dehydrati on, subjects rested for 30 mi n i n a contour
chai r, wi thout access to ui ds; after 30 mi n, they drank water
ad l i bi tum for 180 mi n. Bl ood sampl es (20 ml ) were taken just
before dri nki ng (time 0) and at 30, 60, 120, and 180 mi n of
rehydrati on. Uri ne was col l ected at the end of exerci se and at
hourl y i nterval s duri ng rehydrati on, and the uri ne sampl es
were anal yzed for vol ume and sodi um excreti on.
Bloodsamples. Subjects were semi recumbent duri ng pl ace-
ment of the catheter (21 gauge) and were seated for 60 mi n
before sampl es were taken to ensure a steady state i n pl asma
vol ume and consti tuents. Free-owi ng venous bl ood was
obtai ned for the measurement of hematocri t (Hct), pl asma
osmol al i ty (P
Osm
), PRA, P
[AVP]
, P
[ALD]
, P
[E
2
]
, and pl asma con-
centrati ons of atri al natri ureti c pepti de (P
[ANP]
) and pro-
gesterone (P
[P
4
]
). An al i quot (0.5 ml ) was removed for i mmedi -
ate assessment of Hct i n tri pl i cate by mi crohematocri t.
Second and thi rd al i quots were transferred to a hepari ni zed
tube and a tube wi thout addi ti ve, and al l other al i quots were
pl aced i n tubes that contai ned EDTA. The tubes were centri -
fuged, and the pl asma taken off the hepari ni zed sampl e was
anal yzed for al dosterone. P
[E
2
]
and P
[P4]
were measured by
usi ng serum from the tube wi thout addi ti ve. The EDTA
sampl es were anal yzed for P
[AVP]
, P
[ANP]
, and PRA. Al l bl ood
sampl es were anal yzed for Hct, P
Osm
, P
[ALD]
, P
[AVP]
, P
[ANP]
, and
PRA; onl y the basal bl ood sampl es were al so anal yzed for P
[E
2
]
and P
[P
4
]
.
Blood Analysis
P
Osm
was measured by freezi ng-poi nt depressi on (Ad-
vanced I nstruments 3DI I ); P
[ALD]
, P
[AVP]
, P
[ANP]
, P
[E2]
, and P
[P
4
]
were measured by radi oi mmunoassay. I ntra- and i nterassay
coeffi ci ents of vari ati on for the mi drange standards were,
respecti vel y, as fol l ows: P
[AVP]
(4.52 pg/ml ), 6.0 and 3.4%
[I mmuno Bi ol ogi cal Labor ator i es (I BL), Hambur g, Ger-
many]; PRA (4.5 ng ml
1
ANG h
1
), 2.3 and 2.9% (Di asori n,
Sti l l water, MN); P
[ALD]
(132 pg/ml ), 3.4 and 3.6% (Di agnosti c
Products, Los Angel es, CA); P
[ANP]
(63.3 pg/ml ), 5.1 and 5.2%
(Di asori n); P
[E
2
]
(64.3 pg/ml ), 3.7 and 4.0% (Di agnosti c Prod-
ucts); and P
[P
4
]
(3.7 pg/ml ), 2.1 and 2.5% (Di agnosti c Prod-
ucts). The assay for AVP has a sensi ti vi ty of 0.8 pg/ml ; thi s
sensi ti vi ty i s necessary to detect smal l , but i mportant, changes
i n thi s hormone.
Statistical Analysis
Pearsons product-moment correl ati on on i ndi vi dual data
was used to assess the sl ope and absci ssal i ntercepts of the
P
[AVP]
-P
Osm
rel ati onshi p duri ng dehydrati on (6). The wi thi n-
phase rel i abi l i ty of our most i mportant dependent vari abl es
(ui d-regul ati ng hormones and osmoti c regul ati on of AVP, as
measured at rest, dehydrati on, and rehydrati on) was deter-
mi ned wi th Cronbachs , assumi ng a val ue 0.80 as an
acceptabl e l evel of rel i abi l i ty (2). Areas under the curve (AUC;
trapezoi d method) were cal cul ated duri ng the rehydrati on
peri od (starti ng 30 mi n postexerci se) for PRA, P
[ALD]
, and
P
[ANP]
, and thei r rel i abi l i ty was determi ned wi thi n a gi ven
menstrual cycl e by usi ng Cronbachs . We used repeated
measures ANOVA model s, fol l owed by Bonferroni s t-test to
test di fferences i n the dependent vari abl es both wi thi n and
between menstrual phases. Data were anal yzed by usi ng
BMDP stati sti cal software (BMDP Stati sti cal Software, Los
Angel es, CA) and were expressed as means SE.
RESULTS
Al l subjects were tested duri ng the rst 5 days (4 1
days) after the start of menstrual bl eedi ng for earl y
fol l i cul ar-phase tests, and between 20 and 25 days
(22 2 days) for the mi dl uteal -phase tests. Speci cal l y,
the subjects were tested between days 7 and 10 after
the LH peak, and, therefore, 69 days after ovul ati on.
Between-PhaseMeasurements
At rest, Hct, P
[E
2
]
, P
[P
4
]
, P
[ALD]
, and PRA were hi gher,
and P
Osm
and P
[ANP]
were l ower, i n the l uteal phase
compared wi th the fol l i cul ar phase (P 0.05); however,
there were no di fferences i n body wei ght or P
[AVP]
(Tabl es 1 and 2). Duri ng dehydrati ng exerci se, body
water l oss (1.5 0.2 kg, or 2.3% of preexerci se body
wei ght) was comparabl e between the fol l i cul ar and
mi dl uteal phases. Si mi l arl y, despi te the basel i ne vari -
abi l i ty, P
[AVP]
and PRAresponses to exerci se (i .e., change
from basel i ne) were si mi l ar between the two phases
(Tabl e 2). However, thi s was not the case for P
[ALD]
, i n
whi ch the exerci se response was greater duri ng the
mi dl uteal phase. Li near regressi on anal ysi s of the
i ndi vi dual subjects data duri ng dehydrati on i ndi cated
si gni cant correl ati ons between P
[AVP]
and P
Osm
, wi th r
val ues rangi ng from 0.82 to 0.98. The absci ssal i nter-
cept of the l i near P
[AVP]
-P
Osm
rel ati onshi p, or theoreti -
cal osmoti c threshol d for AVP rel ease, was l ower i n the
mi dl uteal phase (278 1 and 279 1 mosmol /kgH
2
O;
Tabl e 1 and Fi g. 1) compared wi th the fol l i cul ar phase
(282 1 and 283 1 mosmol /kgH
2
O; P 0.05). The
sl opes of thi s rel ati onshi p were unaffected by men-
strual phase. Duri ng rehydrati on, the AUCs for P
[ALD]
and PRA were si gni cantl y greater i n the l uteal com-
pared wi th the fol l i cul ar phase.
Within-PhaseMeasurements
Early follicular phase. Wi thi n the fol l i cul ar phase,
there were no si gni cant di fferences among the means
1093 FLUI D-REGULATI NG HORMONES I N YOUNG WOMEN

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of any of the vari abl es duri ng rest, dehydrati on, and
rehydrati on. However, wi th the excepti on of P
[ANP]
,
none of the resti ng val ues of the ui d-regul ati ng hor-
mones attai ned suffi ci entl y hi gh Cronbachs to be
consi dered rel i abl e (Tabl e 3). Rel i abi l i ty for P
[AVP]
and
PRA was better after dehydrati ng exerci se, al though
rel i abi l i ty remai ned l ow for P
[ALD]
( 0.66) and
remai ned hi gh for P
[ANP]
( 0.90). Duri ng dehydra-
ti on, both the sl ope and absci ssal i ntercept of the
P
Osm
-P
[AVP]
rel ati onshi p were hi ghl y rel i abl e wi thi n the
fol l i cul ar phase, attai ni ng Cronbachs of 0.96 and
0.90, respecti vel y. Agai n, P
[AVP]
, P
[ALD]
, and PRA were
not rel i abl y reproduced duri ng rehydrati on, whereas
Cronbachs for P
[ANP]
was 0.93. P
[E
2
]
was hi ghl y
reproduci bl e wi thi n the fol l i cul ar-phase tests, attai n-
i ng Cronbachs of 0.85, but P
[P
4
]
attai ned a Cronbachs
val ue of onl y 0.62 between tests i n the fol l i cul ar
phase.
Midluteal phase. As i n the fol l i cul ar phase, there
were no di fferences i n mean hormonal concentrati ons
at rest, after dehydrati on, or duri ng rehydrati on wi thi n
the mi dl uteal phase. Agai n, resti ng val ues for P
[AVP]
,
P
[ALD]
, and PRA were not hi ghl y reproduci bl e between
the two mi dl uteal phase tests (Tabl e 3). Rel i abi l i ty for
P
[ANP]
was greater, compared wi th the other ui d-
regul ati ng hormones, at rest and duri ng exerci se and
rehydrati on. Despi te hi gh l evel s of rel i abi l i ty for os-
moti c regul ati on of AVP (Tabl e 3), resti ng and rehydra-
ti on l evel s of P
[AVP]
were not consi stentl y correl ated
wi thi n the l uteal -phase tests. I n contrast to the fol l i cu-
l ar phase, however, both P
[E
2
]
and P
[P
4
]
were hi ghl y
consi stent between the two l uteal -phase tests, yi el di ng
Cronbachs val ues of 0.93 and 0.93, respecti vel y.
DISCUSSION
We exami ned the wi thi n-phase physi ol ogi cal rel i abi l -
i ty of the ui d- and sodi um-regul ati ng hormone concen-
trati ons i n the pl asma over two nonconsecuti ve men-
strual cycl es (1216 wk apart)duri ng the earl y fol l i cul ar
and mi dl uteal phases. P
[AVP]
, P
[ALD]
, and PRA vari ed
wi thi n each of the di fferent menstrual phases; however,
there were no stati sti cal di fferences among the means
of any of these hormone concentrati ons. Thi s i ndi cates
that the wi thi n-subject vari abi l i ty remai ns undetected
when onl y the means are tested or reported. Nonethe-
l ess, our data i ndi cate that between-phase di fferences
i n the hormone concentrati ons far exceed the vari abi l -
i ty wi thi n the phases, and, therefore, the l ow wi thi n-
phase rel i abi l i ty does not prevent the detecti on of
menstrual -phase-rel ated changes i n these vari abl es. I n
contrast, P
[AVP]
, PRA, and P
[ANP]
responses to dehydra-
ti on were hi ghl y rel i abl e wi thi n each menstrual phase;
thi s i ndi cates that hormonal responses to stress are
Tabl e 2. Fluid-regulation hormoneconcentrations
at rest and duringexerciseand rehydration in early
follicular and midluteal phases of menstrual cycle
Preexerci se,
0 mi n
Exerci se,
150 mi n
Rehydrati on,
AUC
Follicular phase
P
[ALD]
, pg/ml
Trial A 7912 27565 22810
2
3710
2
Trial B 9619 19847 16610
2
3010
2
PRA, ng ml
ANG
1
h
1
Trial A 0.80.2 3.91.0 28760
Trial B 0.90.2 3.41.1 26762
P
[AVP]
, pg/ml
Trial A 1.30.2 3.70.8 39972
Trial B 1.20.4 3.50.8 374106
P
[ANP]
, pg/ml
Trial A 33.03.9 88.111.7 7810
2
810
2
Trial B 38.05.3 87.912.1 7610
2
810
2
Luteal phase
P
[ALD]
, pg/ml
Trial A 15722* 38843* 33010
2
4710
2
*
Trial B 15521* 50051* 46010
2
5210
2
*
PRA, ng ml
ANG
1
h
1
Trial A 1.80.4* 6.11.7* 471113*
Trial B 1.70.2* 4.20.9* 653121*
P
[AVP]
, pg/ml
Trial A 1.20.2 3.20.6 34779
Trial B 1.10.3 3.71.1 496125
P
[ANP]
, pg/ml
Trial A 49.65.6 109.214.5 9410
2
910
2
Trial B 54.69.2 114.822.2 10110
2
1410
2
Val ues are means SE. Trials A and B are rst and second tri al s,
respecti vel y, wi thi n speci ed menstrual phase. AUC, area under the
curve (trapezoi d). Pl asma reni n acti vi ty (PRA), P
[AVP]
, and pl asma
concentrati ons of al dosterone (P
[ALD]
) and atri al natri ureti c pepti de
(P
[ANP]
) at rest, and i n response to dehydrati ng exerci se and 180 mi n
of ad l i bi tum rehydrati on i n earl y fol l i cul ar and mi dl uteal phases of
menstrual cycl e. *Si gni cant di fference between fol l i cul ar and l uteal
phases, P 0.05.
Tabl e 1. Subject characteristics in earlyfollicular
and midluteal phases of themenstrual cycle
Characteri sti cs
Fol l i cul ar
Phase
Luteal
Phase
Body wei ght, kg
Trial A 61.73.6 61.33.5
Trial B 61.53.9 61.63.7
Hematocri t, %
Trial A 36.60.8 36.81.0*
Trial B 36.50.7 37.90.9*
P
[E2]
, pg/ml
Trial A 26.94.6 98.916.6*
Trial B 20.74.1 128.120*
P
[P4]
, ng/ml
Trial A 1.30.4 8.72.0*
Trial B 0.90.4 9.82.3*
P
Osm
-P
[AVP]
sl ope, pg ml
1
mosmol
1
Trial A 0.470.11 0.510.18
Trial B 0.490.14 0.560.17
P
Osm
-P
[AVP]
x-i ntercept, mosmol /kgH
2
O
Trial A 2832 2791*
Trial B 2831 2791*
Val ues are means SE; Trial A and Trial B are the rst and second
tri al s, respecti vel y, wi thi n the speci ed menstrual phase. Preexerci se
body wei ght, hematocri t, and pl asma concentrati ons of 17-estradi ol
(P
[E
2
]
) and progesterone (P
[P
4
]
) i n the earl y fol l i cul ar and mi d-
l uteal phases of the menstrual cycl e. Sl opes and absci ssal i ntercepts
are based on i ndi vi dual subjectspl asma argi ni ne vasopressi n concen-
trati on (P
[AVP]
)-pl asma osmol al i ty (P
Osm
)rel ati onshi p duri ng dehydra-
ti on i n the earl y fol l i cul ar and mi dl uteal phases of the menstrual
cycl e. *Si gni cant di fference between fol l i cul ar and l uteal phases,
P 0.05.
1094 FLUI D-REGULATI NG HORMONES I N YOUNG WOMEN

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more consi stent, despi te the vari abi l i ty i n basel i ne
val ues.
Al though there were no si gni cant wi thi n-phase
di fferences between the means of the sodi um-regul at-
i ng hormones, onl y P
[ANP]
val ues were consi stentl y
rel i abl e duri ng rest, exerci se, and rehydrati on wi thi n
ei ther of the two phases. Resti ng P
[AVP]
, P
[ALD]
, and PRA
were qui te vari abl e across the two tri al s wi thi n both
the fol l i cul ar and l uteal menstrual phases. I ndeed, thi s
basel i ne vari abi l i ty exi sts even wi th careful control of
predehydrati on water and sodi um i ntake, posture, and
ti mi ng of the experi ments to coi nci de wi th speci c
events duri ng the menstrual cycl e (such as ovul ati on
and menses). Resti ng or basal vari ati ons i n P
[AVP]
may
be exaggerated further by the fact that val ues were
cl ose to the l owest l evel of sensi ti vi ty of our assay
techni que (i .e., 0.8 pg/ml ). Al so, because the rehydra-
ti on was ad l i bi tum, hydrati on-recovery rates may have
been di fferent among the test days. Therefore, al though
total ui d i ntake was si mi l ar over the four tests,
changes i n dri nki ng patterns or dri nki ng rates may
substanti al l y affect AVP rel ease at a gi ven bl ood sam-
pl i ng poi nt (17) and, consequentl y, affect our abi l i ty to
observe repeatabl e P
[AVP]
.
I n any case, despi te the l ow wi thi n-phase rel i abi l i ty
of P
[AVP]
at rest and duri ng rehydrati on, osmoti c regul a-
ti on of P
[AVP]
(i .e., sl opes and i ntercepts)duri ng dehydra-
ti on was hi ghl y reproduci bl e. Thi s i ndi cates that, al -
though i ndi vi dual val ues may vary, the regul ati on of
thi s hormone i n response to envi ronmental stress (e.g.,
exerci se) remai ns constant. Thi s i s an i mportant nd-
i ng, because smal l shi fts i n the regul ati on of AVP l ead
to l arge changes i n renal water retenti on (13). More-
over, al though a number of studi es have demonstrated
changes i n osmoti c regul ati on of P
[AVP]
over the course
of a si ngl e menstrual cycl e (18, 19), the menstrual -
phase effects on the P
Osm
threshol d for AVP rel ease are
onl y 56 mosmol /kgH
2
O, maki ng essenti al a preci se
and consi stent measurement of the P
Osm
i ntercept
wi thi n a gi ven menstrual phase.
I nteresti ngl y, the shi fts i n osmoti c regul ati on of AVP
and the ui d regul ati on hormones over the course of
the menstrual cycl e do not seem to i mpact overal l body
ui d and sodi um retenti on. Despi te the shi ft i n osmoti c
AVP regul ati on, ui d l oss duri ng exerci se was si mi l ar
i n both menstrual phases. I n the l uteal phase, a
progesterone-i nduced i nhi bi ti on of al dosterone-depen-
dent sodi um reabsorpti on at di stal si tes i n the nephron
causes transi ent natri uresi s (12). Thi s natri uresi s i s
fol l owed by a compensatory sti mul ati on of the reni n-
al dosterone system (9, 16, 20), resul ti ng i n a sl i ght
attenuati on of sodi um excreti on duri ng the l uteal phase
(7.2 1.4 vs. 11.5 2.0 meq). Nonethel ess, overal l
water and sodi um bal ance appear unaffected by the
shi fts i n ei ther progesterone or the sodi um regul ati on
hormones (9). Thi s l eads us to specul ate that estrogen
and progesterone have thei r pri mary i mpact on body
water regul ati on through changes i n body water and
sodi um di stri buti on rather than through retenti on.
We al so tested the rel i abi l i ty of the femal e sex
hormones 17-estradi ol and progesterone. P
[E
2
]
was
hi ghl y reproduci bl e between the two tri al s i n both the
fol l i cul ar and mi dl uteal phases. P
[P
4
]
, al though reproduc-
i bl e duri ng the l uteal phase, was somewhat vari abl e
between the two tri al s i n the fol l i cul ar phase. P
[P
4
]
i s
normal l y l ow duri ng the fol l i cul ar phase of the men-
strual cycl e, so even smal l vari ati ons l ead to l arge error
val ues and may thus exaggerate the vari abi l i ty of P
[P
4
]
duri ng the fol l i cul ar phase. Nonethel ess, despi te the
l ow rel i abi l i ty, P
[P4]
val ues were consi stent and l ow
enough to i ndi cate the subjects were i n the fol l i cul ar
phase of the menstrual cycl e.
The vari abi l i ty i n the ui d-regul ati ng hormones was
not substanti al enough ei ther to create si gni cant
stati sti cal di fferences i n means between tri al s wi thi n
the same menstrual phase or to obscure the l arge
di fferences i n these hormone concentrati ons between
menstrual phases. Nonethel ess, these ndi ngs suggest
that there i s a natural vari abi l i ty i n these hormone
Fi g. 1. Mean pl asma argi ni ne vasopressi n concentrati on (P
[AVP]
)
responses to i ncreases i n pl asma osmol al i ty (P
Osm
) duri ng dehydra-
ti on i n fol l i cul ar and l uteal phase tests. Trials A and B are the rst
and second tri al s, respecti vel y, wi thi n the speci ed menstrual phases.
Data are means SE.
Tabl e 3. Cronbachs for reliabilitywithin 2follicular
and 2luteal phasetests
Cronbachs
Fol l i cul ar phase Luteal phase
Resti ng P
[AVP]
0.49 0.25
Exerci se P
[AVP]
0.81* 0.98*
Rehydrati on P
[AVP]
0.58 0.96*
P
[AVP]
-P
Osm
sl ope 0.96* 0.81*
P
[AVP]
-P
Osm
i ntercept 0.90* 0.86*
Resti ng P
[ANP]
0.80* 0.80*
Exerci se P
[ANP]
0.90* 0.87*
Rehydrati on P
[ANP]
0.93* 0.80*
Resti ng PRA 0.49 0.51
Exerci se PRA 0.72 0.89*
Rehydrati on PRA 0.67 0.95*
Resti ng P
[ALD]
0.55 0.66
Exerci se P
[ALD]
0.66 0.82*
Rehydrati on P
[ALD]
0.64 0.76
Resti ng P
[E
2
]
0.85* 0.93*
Resti ng P
[P
4
]
0.62 0.92*
*Cronbachs 0.80 was consi dered rel i abl e.
1095 FLUI D-REGULATI NG HORMONES I N YOUNG WOMEN

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responses, whi ch may be undetected when onl y grouped
mean val ues are presented.
We grateful l y acknowl edge the techni cal support of Tamara S.
Morocco, John R. Stofan, and Ri chard Wempl e, and the cooperati on of
the vol unteer subjects. We al so thank Lou A. Stephenson for contri bu-
ti ons to the research desi gn.
Thi s work i s supported by the US Army Medi cal Research and
Materi el Command under contract No. DAMD1796-C-6093. The
vi ews, opi ni ons, and/or ndi ngs contai ned i n thi s report are those of
the authors and shoul d not be construed as an offi ci al Department of
the Army posi ti on, pol i cy, or deci si on unl ess so desi gnated by other
documentati on.
I n conduct of research where humans are the subjects, the
i nvesti gators adhered to the pol i ci es regardi ng the protecti on of
human subjects as prescri bed by 45 CFR 46 and 32 CFR 219
(Protecti on of Human Subjects).
Address for repri nt requests and other correspondence: N. S.
Stachenfel d, The John B. Pi erce Laboratory, 290 Congress Ave., New
Haven, CT06519 (E-mai l : nstach@jbpi erce.org).
Recei ved 18 September 1998; accepted i n nal form 23 November
1998.
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