BIOLOGY OF REPRODUCTION 24, 967-976 (1981)

Site of Action of Prolactin in the Suppression of Gonadotropin

Secretion During Lactation in the Rat:
Effect on Pituitary Responsiveness to LHRH’

M. SUSAN SMITH2’3

Department of Physiology,
University of Massachusetts Medical Center,
Worcester, Massachusetts 01605

ABSTRACT

The elevated levels of prolactin associated with lactation contribute significantly to the suppres-
sion of the postcastration increase in gonadotropin secretion during lactation. This study deter-
mines the effects of prolactin on pituitary responsiveness to LHRH in the rat. Blood samples were
collected before administration of LHRH (100 or 500 ng) and at 15, 30, 45, and 60 mm after
treatment with LHRFI. Administration of exogenous prolactin on Days 10 and 11 postpartum to
females nursing two pups had no effect on basal LH or FSH secretion Day 11 or on the LH or FSH
responses after LHRH administration. Whereas ovariectomy on Day 10 resulted in a significant
increase in LH secretion on Day 1 1, administration of exogenous prolactin to ovariectomized lacta-
ting females significantly decreased the postcastration rise in LH without having an effect on the
LH response to LHRH. The effect of endogenous prolactin secretion was studied on Day 10 post-
partum in females nursing eight pups by inhibiting suckling-induced prolactin with CB-154 for 24 h
before LHRH administration. The LH response after LHRH was similar in the presence or absence
of elevated levels of prolactin. However, if the pups were removed for 24 h before administration
of LHRH, both basal LH secretion and the response after LHRH increased to levels observed
during diestrus of the estrous cycle.
The effect of endogenous prolactin secretion on the postcastration rise in LH and FSH was
studied on Day 8 or 17 postpartum (5 days after ovariectomy) in females nursing eight pups. The
results showed that whereas suckling plus endogenous prolactin secretion almost completely
blocked any postcastration rise in LH or FSH, suppression of prolactin in the presence of suckling
led to a very large increase in LH and FSH. In spite of large differences in the basal rate of LH and
FSH secretion, LHRH-induced LH secretion was similar in the presence or absence of prolactin.
These results indicate that in intact lactating females, prolactin has no effect acutely to suppress
the basal rate of gonadotropin secretion or LHRH-induced gonadotropin secretion. In ovariecto-
mized lactating females, the elevated levels of prolactin associated with lactation suppress gonado-
tropin secretion but do not appear to be acting at the level of the pituitary to decrease responsive-
ness to LHRH, and therefore must be acting at the level of the hypothalamus.

INTRODUCTION and FSH secretion after ovariectomy. Recent

studies have demonstrated that elevated levels
The ability of lactation to suppress the post-
of prolactin associated with lactation contrib-
castration rise in gonadotropin secretion is well
ute significantly to the suppression of the post-
known (Hammons et al., 1973; Ford and
castration response (Smith, 1978a) and to the
Melampy, 1973; Smith and Neill, 1977; Smith,
suppression of basal gonadotropin secretion (Lu
1 978a). In the rat, suckling of a large litter is as
et a!., 1976b). In addition, administration of
effective as ovarian steroids in suppressing LH
exogenous prolactin during the latter stages of
lactation or to animals suckling two pups causes
a significant decrease in the postcastration rise
in LH and FSH(Muralidharetal., 1977;Smith,
Accepted February 25, 1981.
1978a). During conditions other than lactation,
Received January 21, 1981.
‘This research was supported by a grant from the experimentally induced hyperprolactinemia can
USPHS, NIH (HD-09442). also greatly decrease LH and FSH secretion in
2Recipient of a Research Career Development both male and female rats (Bartke et a!., 1977;
Award from the USPHS, NIH (HD-00264).
Muralidhar et a!., 1977; Winters and Loriaux,
address for reprint requests: Dr. M. Susan
Smith, Dept. of Physiology, University of Pittsburgh 1978; McNeilly et al., 1980). In some studies,
School of Medicine, Pittsburg, PA 15261. decreased pituitary responsiveness to LHRH

967
968 SMITH

has been demonstrated in the hyperprolactine- tin secretion as described previously (Smith, 1978a).
mic animals (Muralidhar et a!., 1977; Winters Females were treated daily at 0900 h with 0.5 mg
CB-154 s.c., a regimen that completely inhibited
and Loriaux, 1978). These results suggest that
milk secretion in lactating females. To maintain nor-
the elevated levels of prolactin during lactation mal pup growth and a vigorous suckling stimulus,
may be acting at the level of the pituitary to litters were exchanged between control and CB-154-
suppress gonadotropin secretion. Generally, the treated females every 12 h.
lactating rat responds to exogenous LHRH in a LH, FSH, and prolactin concentrations in the
serum were determined using the NIAMDD RIA kits.
manner similar to that of diestrous animals
The limit of detection of the LH assay was 10 ng/ml
(Lu et a!., 1976a; Steger and Peluso, 1978; in terms of the NIAMDD-rat LH-RP-1 standard. The
Smith, 1 978b). However, pituitaries of lactating limit of detection of the FSH assay was 50 ng/mI
rats have a greatly decreased ability to sustain (NIAMD-rat FSH-RP-1) and of the prolactin assay was
3.5 ng/ml (NIAMDD-rat prolactin-RP-1). Statistical
LH release as evidenced by the decreased dura-
analyses of differences between means were deter-
tion of the response to LHRH(Lu eta!., 1976a; mined by analysis of variance or Student’s t test.
Smith, 1978b). Signifiance was assessed at the 0.05 level.
These studies were designed to determine
the site of action of PRL in the suppression of
RESULTS
gonadotropin secretion during lactation by
examining the effects of prolactin on pituitary
Effect of CB-154 on
responsiveness to LHRH. The effects of prolac-
LH Release During the Estrous Cycle
tin on the magnitude and duration of the
response to LHRH were studied in cycling fe- To determine whether the regimen of CB-
males and in intact or ovariectomized females 154 treatment, itself, had an effect on LH
nursing two or eight pups. secretion, animals were treated with 0.5 mg CB-
154 s.c. during estrus and diestrus-1. The ef-
MATERIALS AND METHODS fects of exogenous prolactin on LH secretion
were also determined in animals treated with
Female rats (CD strain, Charles River Labs., Wil-
0.75 mg ovine prolactin twice daily during
mington, MA) were housed under a 12 h diurnal light
cycle (lights on 0600 h). Vaginal smears were ob- estrus and diestrus-1. Pituitary responsiveness
tained 6 days per week, and only those animals show- to 500 ng LHRH was assessed during diestrus-2.
ing two consecutive 4 day estrous cycles were used in The results in Fig. 1 show that CB-154 had no
experiments requiring cycling animals. Pregnant fe-
effect on either basal LH secretion, as deter-
males were placed in maternity cages 2 days before
expected parturition. The day after parturition was mined by 0 mm plasma LH concentrations, or
designated Day 1 postpartum, and litter sizes were ad- on LH release after LHRH stimulation. Plasma
justed to 2 to 8 pups on Day 2 postpartum. Lactating prolactin concentrations averaged <10 ng/ml
females were ovariectomized under ether anesthesia
in the CB-154-treated females compared with
via a midventral abdominal laparotomy.
To determine pituitary responsiveness to synthetic
LHRH, animals were anesthesized with ether between
0900 and 1100 h and an aortic catheter was inserted.
All animals were allowed to recover at least 1 h and
.-. nESTRUS-2
lactating females were returned to their litters. The .---I DESTRUS-2+PRL
females were then injected i.p. with sodium pentobar- 200 DIESTRIJS-2 .ce-154

bital (35 mg/kg). LHRH (Parke-Davis) was thawed

immediately before use and diluted with PBS so that 50-
0.5 ml contained 100 or 500 ng LHRH. After collec- E
tion of a controlblood sample (0 time, 0.5 ml), LHRH
was injected slowly over a 15 sec period via the aortic
I
catheter, and blood samples were collected at 15, 30, -J

45, and 60 mm. An equal volume of heparinized saline 50

(0.5 ml) was used to flush the catheters after each
blood sample was taken. Preliminary studies showed
that administration of saline instead of LHRH resulted 5 30 4 60
in no significant change in LH or FSH secretion after MINUTES AFTER LHRH
0 mm. The processed plasma was frozen until hor-
mone analyses were performed. FIG. 1. LH response to 500 ng of LHRH during
Ovine prolactin (27 lU/mg) was injected s.c. in diestrus-2 of the estrous cycle. Ovine prolactin (0.75
saline at the time of ovariectomy and subsequently mg) was injected s.c. at 0900 and 1800 h on estrus
at 0900 and 1800 h daily for the duration of the and diestrus-1. CB-154 (0.5 mg) was injected s.c. at
particular experiment. CB-1 54 (2.Br--ergocryptine, 0900 h on estrus and diestrus-1. Each point on the
Sandoz) was used to inhibit suckling-induced prolac- graph represents the mean ± SEM of 9-10 animals.
 SUPPRESSION OF GONADOTROPINS DURING LACTATION 969

20-50 ng/ml in the control animals. Similarly, N N ‘00 ‘#{216}’.5

-1- r-*
treatment with exogenous prolactin for 2 days -
0 + +4 +4 +4 +4 +4
had no effect on LH secretion on diestrus-2. ‘0 N

Basal FSH was not affected by either treat- ‘i so o

0’.O N’O W5V5
ment. Furthermore, LHRH did not evoke any
significant increase in FSH secretion during the
60 mm after LHRH administration (data not
shown).
0Or’4 00U5 Os’.5

+4 +4 *1 +1 +1 +4

Pituitary Responsiveness to N - P. - -

LHRH in Females Nursing Two Pups - 0’ ‘0’.’ N N

-0 00 N 1.5 VS

The effect of prolactin treatment on pitui-

tary responsiveness to LHRH in intact females
nursing two pupsis shown in Table 1. Treat- E
ment with exogenous prolactin on Days 10 and . N . -

11 postpartum had no effect on pituitary N * 0’. 0’ 00

N -
responsiveness to LHRH on Day 11. Further- +4 +4 *1 +4 +4 ‘4
N-’ 0’* ‘ON
more, when pituitary responsiveness was tested _ P. 1- . *
- _ ‘.‘fl 00’ 0000
4 h after a single injection of ovine prolac tin on ‘ - - -
04
Day 11, LH secretion was similar in control and
prolactin-treated animals after either 500 or
100 ng of LHRH. In subsequent studies, a
much larger dose of prolactin (3 mg) was in- N N 0’ 1 -‘S 0’.
0’- ‘-‘0 -‘0
jected 4 h before LHRH administration. No , - - - N
+4 +1 +1 +1 +1 +1
effect was observed on either basal LH secre-
0. ‘ON NP. ‘.5-,
tion or on pituitary responsiveness to LHRH 0 ‘0” N 0 -

when compared with control animals. -

E
The effect of ovariectomy (Day 10 post-
>4
partum) on basal LH secretion and pituitary
0. 0
C
responsiveness to LHRH on Day 11 is shown in
Fig. 2. LH concentrations (ng/ml) were elevated ‘0*. N 0’ * ‘0
*1.5 NN N”.5
0
significantly at 0 mm in the ovariectomized ‘‘ + + + +
0 ‘00 N* ,-‘,-, 0’
0
animals (63.4 ± 4.8) compared with intact ani- ,,‘.
mals (27.6 ± 4.6). Prolactin treatment begin- N * N N N
O 0
ning at the time of ovariectomy significantly x C

0
reduced the postcastration rise in LH to 44.6 ±
- >4
3.4. However, in spite of the suppressed post-
1. 55 0
castration response, pituitary responsiveness to
LHRH was not affected by prolactin; ovariec- “6
tomy produced a similar augmentation in LH V 00 00 00 , 0 >5
secretion in both groups of animals when corn- 0 0 0 0 0 0 0
- In VS #{149}.5
In - ,- . 00
0.
pared with intact controls.

Relative Contribution of Suckling . . 0 00 00

0’ 0’

and Prolactin to the Decreased Duration . , 8

#{149}C - IJ
ofResponsetoLHRH - ,

- ‘‘ - - . ‘000
The data in Fig. 3 show the LH response to . . . >4 . >-. v n ‘.5
0. v ‘5 * N N
500 ng of LHRH during Day 10 postpartum in _ : : .
O . 0 ‘6
females nursing eight pups. In control animals, .E
LH increased from 10 ng/ml at 0 mm to reach ‘ E u.
4-

peak concentrations of 143.5 ± 22.3 ng/ml at “a.

15 mm after LHRH and then declined to 47.8 ± , -
o,.. -
o,. -
0 X-
- 0. I.. ,

3.2 ng/ml at 60 mm. When the LH response .. ‘6... ‘6.. ‘5 .0 U 0

0 0 O
during Day 10 postpartum is compared with i- c u o. 0. U 0.
970 SMITH

Effect of Prolactin on Pituitary

.-. INTACT-,.2 PUPS
300
.----. ov+2 Pups
Responsiveness to LHRH after Castration
o----OOV-.-2PUPS+PRL
Females nursing eight pups were ovariecto-
250
mized on Day 3 postpartum and divided into
three groups: 1) plus eight pups [OV(8)J;
E 200
2) plus eight pups and daily treatment with
C
CB-154 to suppress endogenous prolactin secre-
50
I
-J tion [OV(8) + CB-1541 ; and 3) pups removed

100
[OV(0)] . LHRH was administered 5 days later
on Day 8. Rat prolactin concentrations (ng/ml)
in the various groups at 0 mm were as follows:
50
intact (8), 189.7 ± 34.7; OV(8), 147.6 ± 29.7;
1 I I I I OV(8) + CB-154, 5.4 ± 1.2; OV(0), 36.4 ± 7.4.
0
0 5 30 45 60 LH concentrations at 0 mm are shown in Fig.

MINUTES AFTER LHRH 4. OV(8)

The group shows that suckling in the
presence of prolactin almost completely sup-
DAY II POSTPARTUM
FIG. 2. LH response to 500 ng of LHRH during pressed the postcastration rise in LH when
Day 11 postpartum. Ovariectomy was performed on compared with animals with no litters [OV(0)).
Day 10, and prolactin (0.75 mg) was injected at 0900 Suppression of prolactin in the presence of
and 1800 hon Day 10 and at 0900 hon Day 11. Each
suckling [OV(8) + CB-154] led to a significant
point on the graph represents the mean ± SEM of 5
animals.
rise in LH to approximately one-half the level
observed in the OV(0) group. The responses to
LHRH in these same animals are illustrated in
Fig. 5. The results indicate that ovariectomy led
the response during diestrus-2 (Fig. 1), it can be to an increase in pituitary responsiveness in all
seen that the magnitude of the LH response is groups. However, in the suckled groups, pitui-
similar in the two groups whereas the duration
of the response is greatly reduced in the lacta-
ting animals. To assess which factors associated
#{149}-. CONTROLS PUPS
with lactation were responsible for the de-
#{149}----* C8-154, 8 PUPS
creased duration of the LH response to LHRH, 0- -0 CB-I54. PRL, 8 PUPS
DAY 10 POSTPARTUM
lactating rats nursing eight pups were treated .--. PUPS REMOVED 245

either with CB-154 for 24 h to inhibit endogen-

ous prolactin secretion, or with CB-154 plus
exogenous prolactin, or the litters were re-
moved for 24 h. As can be seen in Fig. 3, the
various treatments had no effect on the magni-
tude of the LH response to LHRH at 15 mm. E
However, at 60 mm, the LH response had de- 0l

C
clined significantly in all suckled animals, even
I
if prolactin was inhibited. In contrast, if the -J
pups were removed for 24 h, the duration of
the LH response was maintained for 60 mm as
in the diestrous-2 females. Basal LH levels (0
mm) also increased significantly after removal
of the pups, from <10 ng/ml in control animals
to 24.2 ± 2.9 ng/ml, a value similar to basal LH 0 5 30 45 60
concentrations during diestrus. FSH secretion MINUTES AFTER LHRH
in these animals was not affected by any of the
FIG. 3. LH response to 500 ng of LHRH during
treatments nor by LHRH administration. Rat Day 10 postpartum. CB-154 (0.5 mg) was adminis-
prolactin concentrations were 289.6 ± 49.7 tered at 0900 h on Day 9 and Day 10. Ovine prolactin
(0.75 mg) was administered at 0900 and 1800 h on
ng/ml in the control animals, 7.4 ± 1.2 ng/mI in
Day 9 and at 0900 h on Day 10. Pups were removed
the CB-154-treated animals, 8.4 ± 2.4 ng/ml in
at 0900 h on Day 9 postpartum. Each group consisted
the CB-1 54 plus prolactin-treated animals, and of 5-7 animals. Points on the graph represent the
28.7 ± 4.2 ng/ml after the pups were removed. mean ± SEM.
 SUPPRESSION OF GONADOTROPINS DURING LACTATION 971

OVARIEC1OMY,
These same experiments were repeated
DAY 3 POSTF*RTUM during late lactation when the suppressive ef-
ISO
fects of suckling were beginning to wane.
Groups of animals were ovariectomized on Day

25
12 and injected with LHRH on Day 17 post-
partum. Rat prolactin concentrations (ng/ml)
in the various groups at 0 mm were as follows:
00
E intact (8), 157.4 ± 24.3; OV(8), 101.4 ± 24.6;

C
I OV(8) + PRL, 74.3 ± 22.4; OV(8) + CB-154,
75 5.4 ± 1.2; OV(0), 24.3 ± 3.6. Based on the 0
I
-J
mm LH concentrations shown in Fig. 6, a
50 significant postcastration rise in LH was ob-
served on Day 17 [OV(8)l. Suppression of pro-
lactin secretion in the continued presence of
25
suckling led to a full postcastration response
[OV(8) + CB-1541, whereas administration of
on ___________________ -- - - exogenous prolactin completely prevented any
INTACT(8) OV(8) OV(8)
OV(0) rise in LH secretion [OV(8) + PRL]. However,
+ CB-154
the inverse relationship between prolactin and
DAY8 POSTPARTUM
LH secretion was not manifested in terms of
FIG. 4. LH concentrationsduring Day 8 postpar- pituitary responsiveness to LHRH. As seen in
tum at 0 mm.
Ovariectomy was performed on Day 3 Fig. 7, the magnitude of the response to 500 ng
postpartum and pups were removed lOV(0)I. CB-154
(0.5 mg) was administered at 0900 h daily from Days of LHRH at 15 mm was similar in all groups of
3-8. Litters were exchanged every 12 h between ovariectomized females regardless of the rate of
OV(8) and OV(8) + CB-154 groups to ensure a vigor- LH release at 0 mm. Whereas the presence or
ous suckling stimulus. Each bar represents the mean of
absence of prolactin appeared to have no effect
7 animals and the line on top of the bar represents the
SEM. on the peak LH concentrations at 15 mm, the
presence of prolactin did affect the duration
of the response to LHRH. In the OV(8) +

tary responsiveness was not affected by the

presence [OV(8)] or absence or prolactin #{149}.-.INTACT, 8 PUPS

[OV(8) + CB-154] even though the rate of .-- -.. OV, 8PUPS
0- OV. BPUPS,C8-154
basal LH secretion at 0 mm was affected. The -#{176}

400 OV,OPJPS DAYS POSTPARTUM

suckling stimulus, itself, was responsible for a
significant suppression of pituitary LH release
350 -
and responsiveness to LHRH as indicated by -

the greater release of LH in the OV(0) group

300
than in the OV(8) CB-154 group.
+
/
FSH concentrations in these same animals
25( /
are shown in Table 2. The 0 mm FSH concen-
E /
trations indicate that suckling in the presence
of prolactin completely suppressed the post-
C 200 /
I
-J
castration rise in FSH [OV(8)), whereas inhib-
ISO
ition of prolactin secretion in the presence of
suckling [OV(8) + CB-1541 led to a significant
00
rise in FSH secretion, which approached the
levels observed in the absence of suckling
50
[OV(0)]. After LHRH administration to intact
females, no increase in FSH secretion was ob- I 1 I I I

served. LHRH administration to the ovariec- 0 5 30 45 60

tomized animals resulted in small increases of MINUTES AFTER LHRH

FSH secretion in each group. Therefore, the
FIG. 5. LH concentrations after 500 ng of LHRH
peak FSH response was related directly to the on Day 8 postpartum. These data are from the same
rate of basal FSH secretion at 0 mm. animals shown in Fig. 4.
972 SMITH

InN 0’..en
(25
0’ N In 0
N * N 0’ OVARIECTOMY,
0 +1 +1 +1 +1
‘0 O’...,NP. (00 DAYI2 POSTPARTUM
55 0’. ‘Oen
00 * 0’ N
- N’0- E
0’
75

I
-j

000’
N 554040 INTACT(S) OV(8) OV(8) (8) OVID)
In +1 +1 +1 +1 *PRL +CB-154
* InNNm
In*enO DAY 7 POSTPARTUM
P. * - P.
55 en

0
FIG. 6. LH concentrations during Day 17 postpar-
C tum at 0 mm. Ovariectomy was performed on Day 12
‘5
0.
postpartum and pups were removed [OV(0)I. CB-154
0 or prolactin was administered once or twice daily,
0.
00
respectively, from Days 12-17. Each bar represents
>4
‘5
the mean of 5-7 animals. (See Fig. 4 for additional
0 E details.)
en 55’0..
C
‘0’0 55 N
0
C
0 +1 +1 +1 +1
en
0 x r-0enI

5 * 55 In N
(‘4 N 0’. 0.
C N en N -

C
CB-154 group, LH concentrations remained at
‘5
V
peak levels through 60 mm, which contrasted
0
‘4
1- to the great decrease in LH at 60 mm observed
in the OV(8) group. The greater release of LH
-J from 30 to 60 mm after LHRH in the OV(0)
V 0’. than in the OV(8) + CB-154 group indicates
#{188}. ene4
‘4
In ‘00’. N that the suckling stimulus, itself, had some
(‘140 In

E In +1 +1 +1 +1 additional suppressive effect on pituitary

0t4 N’ONO
C responsiveness to LHRH.
N N en -
(‘4 VS * 0’
C N en 55- As can be seen in Table 3, FSH concentra-
0
V
tions were significantly elevated 5 days after
U
V

In
U.
C 00
#{149}-#{149}
INTACT. 8 PUPS
0 en #{149}‘--.DV, 8 PUPS
0 U,
E >4 #{149}--. OV.8PtPS,PRL DAY IT POST)RTUM
0
‘5 DV. GPUPS,CB-154
e P.
0
0
‘4 O---O

‘4 .--. OV.OPUPS
‘.0 0 ‘0 en 0. E
00 N N N en
C
‘5 0
0
+1 -H +1 +1 .0
CIOON 0.
U
0’ (‘4 (‘4 N 0
0 . >4 0 U
0’. * VS 0
OIl ‘5 0’ .
0
C C
‘4 0. C
0 0’
C C
0
U ‘5
V U V I
‘5 -J

Ii
0 C ,.
. .
0.
4-
V bO ‘4
0
In
o 0.
U
U
1.5 10
In
10 +I

(‘4 U C.-
‘4
10 .cj +. V > MINUTES AFTER LHRH
0.
0
0
44 .0
FIG. 7. LH response to 500 ng LHRH during Day
I-’ .00o 17 postpartum in the same animals shown in Fig. 6.
 SUPPRESSION OF GONADOTROPINS DURING LACTATION 973

Os In 55 ovariectomy (OV(8)]. Whereas administration

en ‘0 CI - of exogenous prolactin caused a complete sup-
en P. VS ‘0-
CI --
pression of
0 +1
P.
+1
ee
+1
VS
+1 +1
N [OV(8) + PRL],the inhibition
postcastration rise
of endogenous
in FSH
pro-
Os N N en - lactin secretion in the continued presence of
suckling led to a near maximum increase in
FSH secretion [OV(8) + CB-1 54]. Administra-
tion of LHRH resulted in small increases in
FSH secretion. Peak FSH responses in the
‘01- N N ovariectomized groups were different and
P-000O55
en P. * Os en directly related to the rate of basal FSH secre-
1.5
*
+1
ens0en*
+1 +1 +1 +1 tion at 0 mm.
0’. 55 NN en
CI *1- 0’ (‘4
C’4*SV1O’-C’.
0 DISCUSSION
C
‘4
These results confirm and extend earlier
studies (Smith, 1978a) demonstrating that the
0’. elevated levels of prolactin associated with
>4
In en 1-0 In lactation play an important role in the suppres-
0 E
N en 0’ sion of the postcastration rise in gonadotro-
- - pins during lactation. The site of action of pro-
C CI H+I+I+IH

en enJ 1- NJ ‘. lactin in this suppressmon does not appear to be

‘4 00VS-,N1-
P. - 55 * * the pituitary and, therefore, may likely be at
‘-‘*1-NO’
4-
‘4 the hypothalamus. Whether prolactin may be
V affecting hypothalamic function directly or
- .

mndirectly by its effects on other parts of the

CNS is not known at this time.
The data from intact females nursing two
‘0 N In N ‘4. pups demonstrate that administration of exo-
‘4
* Os CI P. genous prolactin had no effect on basal gona-
en en
VS +1 +1 +1 +1 41 dotropin secretion or on the LH response to
.505 en VS -I en 0’ LHRH (Table 1). Conversely, inhibition of
0’. en N Os
o Os’055N0
.9 ‘ 1- en N CI endogenous prolactin secretion for 24 h in
“4 N females nursing eight pups did not restore basal
en
U I LH secretion or the duration of the LH re-
N
en
U, . sponse to LHRI-I to the levels observed in di-
estrous females (Figs. 1, 3). However, removal
C 01
of the pups for 24 h did increase LH secretion
.9 E 0 - significantly. These results suggest that it is the
0 ‘4

N ‘1en CI ‘4. N
‘0 CI 0. CI 0 stimulus,
sucklingsuppression
the of itself,
basal that
LH issecretion
responsible duringfor
enenNIn0’. CI
00 “

-H +1 +1 +1 +1 0 0. >4 lactation. Lu et al. (1976b) found that prolac-

en0’,P.*Q’.
U CI P. ‘4 CI a tin secretion had to be suppressed for several
I 0 P. 55 CI days during lactation before basal LH secretion
N * .-I N 0’.
0. increased. Thus, prolactin may play a more

0C #{176} secondary role in that it is required for the

U C V
‘5 ‘5 V suppressive effects of suckling to be maintained
0 .5 for a long time. However, prolactin does not
4-
0.

0
#{188}.
0
‘I-
V 04 appear to exert an acute effect on gonadotro-
0
U ‘ S VS pin secretion in intact females.
U 10 In 0
#{188}.. ,. In t. In nonlactating animals, numerous studies
. . have been able to demonstrate that experimen-
++ ‘5 ‘4 - ‘ 0.
10 o V V tally induced hyperprolactmnemia can inhibit
0.
O0.L)o.
‘5 .0 U ‘0 u basal gonadotropin secretion in intact rats
i .5 o 0 o o (Bartke et aL, 1977; Hodson et al., 1980;
974 SMITH

McNeilly et al., 1980; Vasquez et al., 1980a,b). possible that the mechanism of prolactin action
However, on an acute basis, prolactin had little during lactation differs from that of other non-
effect (Celotti et al., 1978; Winters and lactating states. Earlier evidence (Smith, 1978a;
Loriaux, 1978). In some studies, pituitary Muralidhar et a!., 1977) showed that a regimen
responsiveness to LHRH was found to be de- of exogenous prolactmn capable to decreasing
creased (Winters and Loriaux, 1978; Greeley LH and FSH secretion in ovariectomized lacta-
and Kizer, 1979; Vasquez et al., 1980a,b). ting rats had no effect if the pups were re-
Vasquez et al. (1980a,b) found that hyper- moved. These results suggest that suckling is
prolactinemia of 6 days duration in intact permissive to the action of prolactin.
cycling females could suppress both basal gona- One of the surprising aspects of this study
dotropin secretion and pituitary sensitivity to was the difference in LH and FSH responses
LHRH. However, they concluded that the to LHRH administration. With this method of
effects were indirect and due to increased pro- LHRH administration in vivo, FSH secretion
gesterone secretion (Vasquez et al., 1980a). does not increase in any intact animals, male or
It remains to be determined why prolactin can- female. In contrast, if pituitaries from intact
not acutely suppress gonadotropin secretion in animals were incubated in vitro, a significant
intact animals. The acute effectiveness of pro- FSH response to LHRH was observed within 1
lactin in ovariectomized but not intact rats h (Smith and Morello, 1980). An increase in
suggests that the presence of ovarian steroids FSH secretion after LHRH administration in
may interfere with the inhibitory action of vivo was observed in ovariectomized females
prolactin. (Tables 2, 3) and in castrated males (unpub-
The ability of prolactin to suppress acutely lished observations) although the magnitude of
(within 24 h) or chronically (5 days) the post- the responses was quite small. These observa-
castration rise in gonadotropins is clearly tions suggest that gonadal steroids or “folliculo-
demonstrated in this study (Figs. 2, 6, Table statin” (inhibin, Campbell and Schwartz, 1979;
3) and confirms earlier studies (Muralidhar et Rush and Lipner, 1979; Hermans et al., 1980)
al., 1977; Smith 1978a). This effect of pro- may be suppressing pituitary release of FSH in
lactin also has been demonstrated repeatedly response to LHRH. It is also possible that a
in nonlactating animals (Winters and Loriaux, regimen of LHRH administration in vivo that is
1977; Celotti et al., 1979; Greeley and Kizer, appropriate for LH secretion is inappropriate
1979; Hodson et al., 1980; McNeilly et al., for stimulating FSH secretion.
1980; Vasquez et al., 1980a,b). Whereas the The results in Tables 2 and 3 show very
present study clearly shows that prolactin has clearly that prolactin is the most important fac-
no effect on pituitary responsiveness to LHRH, tor responsible for the suppression of FSH
the results from other studies are inconsistent secretion after ovariectomy. It is interesting
in that some showed a decreased pituitary that suppression of the postcastration rise in
responsiveness due to prolactin (Celotti et al., FSH is the only aspect of FSH secretion that is
1978; Greeley and Kizer, 1979) whereas others decreased by lactation, since basal FSH secre-
showed no effect of prolactin (Vasquez et al., tion in intact lactating females is normal (Smith
1980a,b). There are several reasons that could and Neill, 1977) (Tables 2, 3) and estrogen-
account for these apparent discrepancies. First, induced FSH surges are similar in magnitude to
the pattern of prolactin secretion and the levels the proestrous FSH surge (Smith, unpublished;
achieved differ greatly in the various studies Coppings and McCann, 1979). In contrast, all
depending on the method used to induce hyper- aspects of LH secretion appear to be suppressed
prolactinemia. Use of pituitary tumors or during lactation. These data suggest that the
pituitary transplants results in chronically suckling stimulus, itself, has little suppressive
elevated levels of prolactin. Several investigators effect on FSH secretion and that the elevated
have demonstrated a correlarjn between the levels of prolactin are primarily responsible for
levels of prolactin achieved and the degree of any inhibition of FSH. Since the effects of pro-
suppression of LH and FSH and the response to lactin appear to be minimal in intact lactating
LHRH (Celotti et al., 1978; Hodson et a!., rats, an effect of prolactin on FSH would be
1980; Vasquez et aI., 1980a,b). During lacta- expected only after ovariectomy.
tion, the pattern of prolactin secretion consists As can be seen in Tables 2 and 3, the incre-
of suckling-induced surges with relatively low ments in FSH secretion after LHRH administra-
levels of prolactin between surges. It is also tion to the ovariectomized females were of
 SUPPRESSION OF GONADOTROPINS DURING LACTATION 975

similar magnitude, regardless of the basal rate adequate pool of LH in a releasable form.
of FSH secretion at 0 mm. Whether FSH con- Pickering and Fink (1979) found that in cycling
centrations were 100-200 ng/ml or 900-1000 rats, a 20-fold increase in the size of the releas-
ng/ml at 0 mm, the maximum increase in FSH able pooi of LH occurred between the morning
was “v150-300 ng/ml. Therefore, the ma.xi- of diestrus and the evening of proestrus, in the
mum FSH concentrations observed after LHRH absence of any significant change in total
differed among the ovariectomized groups and pituitary content of LH. These data suggest
were directly related to the level of FSH secre- that the effects of steroids and LHRH self-
tion at 0 mm. From these data it is difficult to priming to increase the “sensitivity” of the
determine whether prolactin is acting directly pituitary to LHRH may reflect a change in LH
on the pituitary. On the one hand, the similar from a storage form into a readily releasable
increments in FSH secretion after LHRH in the form. If LHRH secretion is significantly
presence or absence of prolactin or suckling decreased during lactation, a hypothesis con-
suggest that prolactin had no effect on pituitary sistent with available data, the ability to main-
responsiveness to LHRH. On the other hand, if tain an adequate amount of LH in a releasable
prolactin has no effect, the peak FSH responses form in the pituitary may be greatly decreased.
after LHRH should be similar in the ovariecto-
mized groups and independent of the rate of ACKNOWLEDGMENTS
FSH secretion at 0 mm. Further studies on the
I would like to acknowledge the expert assistance
regulation of FSH secretion are needed to
of Sharon Alex and Kathryn Morency. Radioimmuno-
resolve this issue. assays for LH, FSH, and prolactin were performed
The LH data from these same animals (Figs. using reagents supplied by the NIAMDD. Ovine pro-
5, 7) are clear and demonstrate that prolactin is lactin also was supplied by the NIAMDD.

not acting on the pituitary to suppress LI-I

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