A possible dual regulation of prolactin release by the serotoninergic system in rats at pro-oestrus and during late pregnancy: role of ovarian hormones

The effect of para-chlorophenylalanine (pCPA), an inhibitor of serotonin synthesis, on prolactin release was studied in rats on the day of pro-oestrus and at the end of pregnancy (day 19). The surges of prolactin normally seen in the afternoon of pro-oestrus in intact rats and in rats ovariectomized on dioestrous day 2 and primed with oestrogen were significantly inhibited by pCPA treatment. Administration of 5-hydroxytryptophan reversed the inhibitory action of pCPA on prolactin release. Treatment with progesterone also completely reversed the inhibitory effect of pCPA on prolactin release in pro-oestrous rats and partially reversed it in ovariectomized oestrogen-treated rats. Ovariectomy on day 19 of pregnancy induced a significant release of prolactin 12 and 24 h later. Administration of pCPA on day 18 of pregnancy produced a marked increase in serum concentrations of prolactin on days 19 and 20 in rats left intact or ovariectomized on day 19. Administration of 5-hydroxytryptophan significantly reversed this stimulatory effect of pCPA on prolactin release but did not modify the release of prolactin induced by ovariectomy. Methiothepin (1-[10,11-dihydro-8-(methylthio) less than b,f greater than thiepin-10,41]-4-methylpiperazine maleate), a serotonin receptor blocker, also induced a significant increase in serum concentrations of prolactin on day 20 of pregnancy in rats left intact or ovariectomized on day 19. These results suggest the existence of different serotoninergic actions in the regulation of prolactin release at pro-oestrus and in late pregnancy. Serotonin facilitates the surges of serum prolactin released at pro-oestrus and in ovariectomized rats treated with oestrogen; progesterone enhances this effect.(ABSTRACT TRUNCATED AT 250 WORDS)     

Inhibitory effect of progesterone on the lactogenic and abortive action of prostaglandin F2alpha.

The effect of ovariectomy, progesterone and prolactin treatment on the action of prostaglandin F2alpha (PGF2alpha) was determined in pregnant rats. PGF2alpha (150 mug times 2) injected i.p. on day 1. or 18 of pregnancy induced lactogenesis about 25 h later and abortion on days 20 and 21 of pregnancy. Treatment with PGF2alpha (100 mug times 2 or 50 mug times 2) on day 19 induced lactogenesis around 22 or 38 h later, respectively, and abortion on day 21. PHF2alpha treatment on day 17 was less effective. Unilateral ovariectomy on day 17 of pregnancy induced lactogenesis 32 h later but not abortion. PGF2alpha (150 mug times 2) given on the day of surgery advanced lactogenesis 12 h and rats aborted on day 19. Bilateral overiectomy on day 17 induced abortion between days 20 to 21, but if a single dose of PGF2alpha (300 mug) was injected on day 18. all the ovariectomized rats aborted on day 19. Progesterone (10 mg) injected into rats treated with PGF2alpha (150 mug times 2) on day 18, prevented abortion and delayed lactogenesis. Prolactin (1 mg times 4) treatment delayed only abortion. Serum prolactin levels were significantly higher 12 h after the last dose of PGF2alpha (150 mug times 2) in rats treated on days 17, 18 or 19 of pregnancy. Pretreatment with progesterone prevented the rise in prolactin concentration. These result suggest that the lactogenic and abortive action of PGF2alpha may be dependent on the uterine and plasma concentration of progesterone.     

Role of the placenta in the control of the ante-partum surge of prolactin in the rat

A nocturnal surge of prolactin secretion occurs in the dark period preceding parturition in the rat. The aim of this study was to examine the role of the placenta in the control of this prolactin surge. Plasma prolactin and progesterone were measured by radioimmunoassay in serial blood samples collected after surgical removal of conceptuses during late pregnancy, and after intracerebroventricular (i.c.v.) injection of placental lactogen (PL) before the prolactin surge. In intact control animals, prolactin secretion remained low until a nocturnal surge of secretion occurred in the dark period preceding parturition, peaking at 269 +/- 51 (S.E.M.) micrograms/l at 03.00 h on day 21. Progesterone levels fell from greater than 200 nmol/l on day 19 to less than 40 nmol/l by 12.00 h on day 20 of pregnancy. PL levels during late pregnancy were modified by partial or complete removal of conceptuses at 10.00 h on day 19 of pregnancy. Removal of all but one or two conceptuses did not change the normal pattern of prolactin or progesterone secretion. Removal of all conceptuses, however, induced a large nocturnal surge of prolactin secretion, peaking at 211.7 +/- 78 micrograms/l at 03.00 h on day 20, 24 h earlier than the surge in intact animals. Progesterone levels after removal of all conceptuses fell to less than 40 nmol/l by 23.00 h on day 19, approximately 12 h before the decline in intact animals. Maintenance of increased progesterone levels after conceptus removal using silicone tubing implants significantly (P less than 0.05) reduced the peak of the premature prolactin surge to 79.7 +/- 18 micrograms/l at 05.00 h on day 20.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effects of pinealectomy, anosmia and blinding on serum and pituitary prolactin in intact and castrated male rats.

Serum prolactin was measured in lightly etherized rats between 4 and 5 a.m. and between 11 a.m. and 12 noon at various times after operation in intact rats and rats that had been operated on. Serum prolactin was elevated in the late nocturnal samples from intact rats and this elevation was prevented by pinealectomy. With time following the operation, daytime serum prolactin was also lowered in pinealectomized animals. In animals rendered anosmic by olfactory bulb removal, serum prolactin was also lowered. Superimposed pinealectomy only lowered late nocturnal serum prolactin in anosmic rats. Blinding the rats by ocular enucleation lowered nocturnal serum prolactin initially and dampened the daily rhythm in prolactin, but superimposed pinealectomy had no effect on the blinded animals. The combined sensory deprivation produced by both blinding and olfactory bulb removal resulted in low prolactin titers regardless of whether blood samples were taken during the day or late at night and, as in the case of blinded animals, there was no effect of pinealectomy. Superior cervical ganglionectomy also lowered serum prolactin. Serum prolactin declined following castration in intact animals and this decline was accentuated by pinealectomy. Either blinding or anosmia also lowered serum prolactin in the castrated animals but, as in intact animals, pinealectomy had little effect on the sensory-deprived rats. A further lowering of prolactin occurred in castrated animals which were both blinded and anosmic; in these animals, pinealectomy again had no additional effect. Measurement of the pituitary prolactin content in the castrated animals at sacrifice revealed a significant lowering of values in blinded animals, with restoration of the values to control levels by pinealectomy. Anosmia also lowered the pituitary prolactin content but this was not reversed by pinealectomy. The lowest pituitary prolactin was found in blinded, anosmic animals and these changes were not reversed by pinealectomy. The weight of the pineal gland increased significantly in anosmic, blinded, or blinded and anosmic rats. It is concluded that pineal secretions in the male rat elevate serum prolactin, particularly late at night, and that loss of input from olfactory and to a lesser extent from visual pathways results in a decrease in the titer of the hormone. This decrease is especially apparent in blinded, anosmic rats.     

Regulation by endogenous opioids of suckling-induced prolactin secretion in pregnant and lactating rats: role of ovarian steroids

Evidence suggests that endogenous opioid peptides are implicated in the suckling-induced prolactin rise. We explored the role of the opioid system and the participation of ovarian hormones in the regulation of prolactin induced by the suckling stimulus at the end of pregnancy in rats with developed maternal behavior, and during lactation. Suckling for 24 h induced a significant increase in serum prolactin on day 19 of pregnancy, which was increased more than three times when naloxone (2 mg/kg s.c.) or mifepristone (2 mg/kg) was administered. The combination of naloxone and mifepristone did not increase serum prolactin more than either compound alone. Administration of tamoxifen (500 microg/kg orally) on days 14 and 15 of pregnancy completely abolished the effect of naloxone, indicating a role for estrogens in establishing this inhibitory role of opioids. To examine the participation of the opioid system during lactation, we used groups of rats on days 1, 3, 5, 12 and 19 postpartum either (i) isolated from the pups for 4 h, or (ii) isolated from the pups for 3.5 h and reunited with them and suckled for 30 min. Naloxone, given just before replacing the pups, prevented the increase in serum prolactin levels observed in the suckled group of rats but had no effect on the basal levels of the isolated rats. To examine whether the participation of the opioid system in the release of prolactin is dependent on the variation of progesterone levels, rats on day 20 of pregnancy were implanted with two cannulae containing progesterone (that blocked postpartum ovulation) or cholesterol, and cesarean surgery was performed on day 21. To maintain lactation, pups (1-3 days old) were replaced every 24 h, and 4 days after the cesarean eight pups were placed in the cage at 1800 h to maintain a strong suckling stimulus during the following 24 h. Naloxone administration significantly reduced serum prolactin levels in control (cholesterol) rats but progesterone implants prevented the inhibitory effect of naloxone and this effect was not modified by treatment with estrogen. These results indicate that the opioid system modulates suckling-induced prolactin secretion, passing from an inhibitory action before delivery to a stimulatory action during lactation. This regulatory shift seems to be dependent on the fall in progesterone concentration at the end of pregnancy and the subsequent increase after the postpartum ovulation and luteal phase.     

Control of corpus luteum function in the pregnant rabbit: role of estrogen and lack of a direct luteotropic role of the placenta

The corpus luteum is essential for pregnancy maintenance in the rabbit and appears to require two luteotropins: estrogen from ovarian follicles and a placental luteotropic factor. We have investigated the role of the placental luteotropic factor in maintaining corpus luteum function in the pregnant rabbit in the absence of estrogen. In Exp 1, follicular estrogen was withdrawn on day 21 of pregnancy by ovulating follicles with 10 IU hCG. In Exp 2, estrogen was withdrawn in hypophysectomized pregnant rabbits on day 21 by removing an estradiol (E2) implant. In the presence of this estrogen implant, luteal function and pregnancy are maintained after hypophysectomy, performed on day 4 of pregnancy. In both experiments, fetoplacental viability was ensured by treating the rabbits with medroxyprogesterone acetate (MPA). In both Exp 1 and 2, withdrawal of estrogen on day 21 of pregnancy caused a dramatic decline in serum progesterone concentrations by day 22. Serum progesterone concentrations remained low, and corpora lutea regressed, although viable fetuses were maintained with MPA. In animals not receiving MPA, estrogen withdrawal caused the loss of luteal function, followed by abortion on days 23-24. In contrast, estrogen replacement (via E2 implant) on day 22 in Exp 1 was fully capable of restoring serum progesterone concentrations to pretreatment values on days 24-27 in MPA-treated rabbits. In rabbits not receiving MPA, estrogen replacement also restored serum progesterone concentrations and prevented abortion. These results provide further evidence that estrogen is essential for normal luteal function in the pregnant rabbit. In the absence of estrogen, the rabbit placenta maintained by the progestagen MPA has no direct luteotropic activity.     

The source of relaxin in pregnant Syrian hamsters

Serum immunoreactive relaxin (IR) was measured on days 8, 10, and 14 of gestation in intact and ovariectomized (day 8 of pregnancy) hamsters. In intact hamsters, IR increased from 3-4 ng/ml on day 8 to 20 ng/ml by day 14 of pregnancy. After ovariectomy on day 8, pregnancy failed, and IR decreased rapidly to 0.29 ng/ml on day 14. However, when pregnancy was maintained in ovariectomized hamsters by daily injections of 0.1 microgram 17 beta-estradiol and 4 mg progesterone, serum IR rose to levels similar to those in intact hamsters on days 10 and 14 of pregnancy (i.e. 15 and 20 ng/ml, respectively). Placentas were obtained from other groups of hamsters on days 11, 14, and 15 of pregnancy and homogenized for bioassay by the classical guinea pig pubic symphysis palpation bioassay. Homogenates of placentas obtained on days 14 and 15 contained, respectively, 4 and 10 micrograms eq porcine relaxin/serum relaxin/g fresh tissue. The placenta, rather than the ovary, appears to be the source of during pregnancy in the hamster.     

The role of prolactin in the luteotrophic process of lactating rats.

Treatment of pseudopregnant rats with ergocryptine mesylate (ECR) depressed serum prolactin levels but had no apparent effect on LH secretion. Ovarian progesterone secretion was significantly, reduced 24 hr after ECR treatment on day 6 or 9 of pseudopregnancy and the secretion rate of 20alpha-OH-P remained constant. When lactating rats nursing 6 pups were treated with ECR on day 6 or 9 postpartum, progesterone secretion was significantly decreased by 48 hr after treatment and 20 alpha-OH-P secretion was not altered. Furthermore, ECR inhibited litter weight gains of these lactating dams. After treatment of normal pregnant and pregnant lactating rats with ECR on day 6 of pregnancy, gestation was terminated in all animals. If ECR was given on day 9 to normal pregnant rats, to pregnant lactating animals whose pups were removed on day 9 postpartum, or to pregnant lactating rats treated with LH, gestation was not terminated. However, treatment with ECR of day 9 pregnant lactating rats whose pups continued to suckle terminated pregnancy in 13 of 21 animals. The results of these studies suggest the elevated pituitary prolactin secretion is necessary for maintenance of luteal function in pseudopregnant and lactating rats, and in pregnant lactating rats elevated pituitary prolactin secretion is a component of the luteotrophic complex for a longer period of time than in normal pregnant rats.     

Failure of rat placenta to inhibit prolactin secretion by ectopic pituitary gland

At mid-pregnancy in the rat, episodic secretion of pituitary prolactin ceases when the placenta is sufficiently developed. At this time, sufficient placental lactogen is secreted by the placenta to inhibit prolactin secretion. The present study tested whether the fully developed placenta at mid-pregnancy can inhibit prolactin secreted by a donor pituitary gland implanted under the kidney capsule. Three pituitary glands were implanted in rats on day 7 of pregnancy; muscle fragments were implanted in controls. Blood was collected during the first and second halves of pregnancy. It was found that prolactin concentrations in the animals with the pituitary implants were high on days 9 and 10 of pregnancy and remained raised during the second half of pregnancy while in control animals nocturnal surges were absent in the second half of pregnancy, the last one occurring on day 10. This observation indicates that the placental hormone cannot act directly on the pituitary gland to inhibit prolactin secretion, but presumably exerts its suppressive effect on prolactin secretion through the hypothalamus.     

Inhibitory effect of the uterus on plasma and pituitary FSH in rats

Plasma concentrations of FSH and LH were measured in ovariectomized, ovohysterectomized, hysterectomized and sham-operated adult, non-pregnant rats at 3, 14, 21 and 28 days after operation. From day 21 after the operation onwards, there were higher concentrations of FSH in plasma in ovohysterectomized than in ovariectomized animals. The concentration of LH was not influenced by hysterectomy. The inhibitory response of FSH and LH to a single dose of oestradiol was not altered by any of the operations. By 2 weeks after surgery, pituitary FSH content had increased in ovohysterectomized animals compared with ovariectomized ones, but this difference was eliminated when ovohysterectomized animals were treated with crude uterine extract. Pituitary contents of LH and prolactin were not influenced by hysterectomy or by treatment with uterine extract, thus indicating the specificity of an inhibitory effect of the uterus on FSH levels. Treatment of hysterectomized and intact animals with uterine extract resulted in a reduction in the weight of the ovaries of 23-38% (P less than 0.05), indirectly showing the presence of an FSH-inhibiting substance in the extract. Fractionated uterine extract inhibited FSH synthesis by rat pituitary cells in vitro, but had no effect on LH synthesis. Chromatographic analysis indicated that the FSH-inhibiting substance in the uterus has a molecular weight of 10,000-20,000.     

Effects of passive immunization against vasoactive intestinal peptide on serum prolactin and LH levels

Recent findings suggest that vasoactive intestinal peptide (VIP) may be a physiological regulator of prolactin secretion and may also be involved in the control of LH secretion. In the present work we have studied the effect of the blockade of endogenous VIP by means of the injection of a specific rabbit anti-VIP serum, in male and female rats with hyperprolactinemia. The administration of 0.5 ml of the VIP antiserum in ovariectomized rats given an acute or chronic treatment with 17 beta-estradiol induced a significant decrease in serum prolactin and LH levels as compared with estrogenized-control rats injected with normal rabbit serum. Anti-VIP serum also reduced serum LH levels in ovariectomized rats not treated with estrogens. The administration of the same antiserum decreased serum prolactin levels in male rats implanted with 2 anterior pituitary glands under the kidney capsule. On the other hand, the injection of the anti-VIP serum in the morning in proestrus rats brought about an increase in serum prolactin and LH levels in the afternoon of the same day. These results confirm previous data showing that VIP is a stimulator of prolactin release, and may also participate in the control of LH secretion in ovariectomized rats acting as a facilitatory factor. During proestrus however, VIP may act in an opposite way, inhibiting, rather than stimulating, prolactin and LH release.     

Regulation of the hepatic growth hormone receptor and serum growth hormone-binding protein during pregnancy in the mouse: effects of litter size.

The regulation of hepatic GH receptor (GHR) and serum GH-binding protein (GHBP) during pregnancy in the mouse was investigated by manipulating the number of conceptuses carried by the dam. Animals carrying 1-4 conceptuses had significantly lower amounts of hepatic GHR (GH-binding activity) than mice carrying 10-13 conceptuses on days 9 and 13 of pregnancy. There was no significant difference in hepatic GHR on day 17 of pregnancy between animals carrying 1-4 and 10-13 conceptuses. Animals carrying 1-4 conceptuses had significantly lower concentrations of GHBP in serum than animals carrying 10-13 conceptuses on days 9, 13, and 17 of pregnancy. The relative amounts of liver GHR- and GHBP-encoding messages in animals with low and high conceptus numbers were investigated by Northern analysis. There were higher levels of both messages in animals carrying 10-13 conceptuses than in mice carrying 1-4 conceptuses. On day 13 of pregnancy, animals carrying 10-13 conceptuses had significantly higher levels of GHBP-encoding message than animals carrying 1-4 conceptuses. Total hepatic mass was not significantly different between animals with low and high conceptus numbers. No significant difference was found in GHBP concentration between blood from the uterine vein and the trunk in 17-day pregnant animals. Mouse placental lactogen-I (mPL-I), mPL-II, GH, and corticosterone concentrations were measured by RIA and related to hepatic GHR activity and serum GHBP concentration. Hepatic GHR activity and serum GHBP concentration were significantly correlated with each other on days 9, 13, and 17 of pregnancy. Hepatic GHR activity was significantly correlated with mPL-I and mPL-II on day 9 of pregnancy. GHBP concentration was significantly correlated with mPL-I and mPL-II on day 9 of pregnancy and with mPL-II and GH on day 13 of pregnancy. Data are consistent with the hypothesis that mPL-I, mPL-II, and GH may affect hepatic expression of the GHR/GHBP gene during pregnancy in the mouse.     

Histoenzymological studies of 3beta- and 17beta-hydroxysteroid dehydrogenases in the placentae and adrenal glands of bilaterally ovariectomized rats

The distribution and activities of 5-unsaturated-3beta- and 17beta-hydroxysteroid dehydrogenases (HSD) have been studied during the pregnancy of normal and ovariectomized rats on days 17, 19 and 21 post coitum. No hormonal substitution was provided after bilateral ovariectomy on day 15 post-coitum. 5-Unsaturated-3beta-HSD was characterized with pregnenolone, 17alpha-hydroxypregnenolone and dehydroepiandrosterone as substrates; oestradiol-17beta and testosterone were used for the investigation of 17beta-HSD activity. With these substrates, it was found that the placental and adrenal activities and distribution of 5-unsaturated-3beta- and 17beta-HSD did not differ in ovariectomized and control rats and it is suggested that increased placental or adrenal steroidogenesis does not supplement deficient ovarian function in order to maintain pregnancy. In the pregnant rat, the ovaries do not control the activities of 5-unsaturated-3beta- and 17beta-HSD in the placenta.     

Further evidence on the role of the hypothalamic afferents on the estrogen-induced prolactin release.

Serum prolactin (Prl) concentrations in ovariectomized rats were low without significant differences between morning and afternoon values. These levels were not affected by either frontal or caudal hypothalamic deafferentation. However, they increased after lesioning the hypothalamic median eminence (ME). Three days after the injection of 20 microgram estradiol benzoate (EB) into ovariectomized non-lesioned rats, a rise in serum Prl occurred in the afternoon but not in the morning. In animals with ME lesions estrogen enhanced both morning and afternoon values. The animals with caudal hypothalamic deafferentation and those which had undergone sham operation showed the same pattern as the normal animals. On the contrary, after estrogen treatment of rats with frontal hypothalamic deafferentation high serum Prl concentration during the morning and low levels in the afternoon were observed. It is concluded that estrogen effects on Prl secretion are in part mediated by frontal neural afferents to the hypothalamus. They would facilitate Prl inhibiting factor (PIF) secretion in the morning and inhibit PIF secretion in the afternoon.     

Ovariectomy leads to a rapid increase in rat placental lactogen secretion

Serum placental lactogen (rPL) levels were measured by RIA in late pregnant rats subjected to a number of endocrine ablations. Adrenalectomy or unilateral ovariectomy had no significant effect on serum rPL levels. Hypophysectomy of the dam at midpregnancy resulted in a significant increase in serum rPL at late pregnancy. Bilateral ovariectomy of day-14 or -16 pregnant rats led to a rapid increase in serum rPL levels for 2 days followed by a gradual decrease as fetuses were reabsorbed or aborted. Adrenalectomy combined with ovariectomy led to sustained, elevated levels of serum rPL which were greater than those seen with bilateral ovariectomy alone. When progesterone (4 mg/rat X day) and estrone (500 ng/rat X day) or 17 beta-estradiol (100 or 200 ng/rat X day) were administered to ovariectomized pregnant rats, serum rPL remained elevated and the conceptuses were retained. In conclusion our studies have shown that ovarian and adrenal factors influence rPL secretion.     

Development and amplification of mid-afternoon surges of prolactin secretion in ovariectomized immature rats

The immature female rat shows a mid-afternoon surge of prolactin secretion which reaches a maximum on the day of first pro-oestrus. The present experiments were undertaken to elucidate the mechanisms which underly the development of this prolactin discharge. Detailed plasma prolactin profiles were obtained from short-term (48 h) ovariectomized rats at 23, 28 or 37 days of age. In the two older groups, but not the youngest, a mid-afternoon surge of prolactin secretion occurred in spite of the absence of the ovaries. To exclude the possibility that such an apparent ovarian-independent discharge of prolactin was due to an oestradiol effect which persisted for 2 days following ovariectomy, another study was conducted using long-term ovariectomized animals. Plasma profiles were obtained from neonatally ovariectomized rats at ages equivalent to juvenile (26-28 days), peripubertal (38-41 days) or adult (46-49 days) phases of development. A mid-afternoon surge of prolactin secretion was observed in the majority of animals (eight out of twelve) irrespective of the interval after ovariectomy; this finding further indicates that in the female rat there is a centrally originated mid-afternoon episode of prolactin secretion which is expressed during juvenile development even in the absence of the ovaries. The relatively small magnitude of these ovarian-independent prolactin discharges (c.f. the preovulatory prolactin surge) suggested that in the intact animal they are amplified by ovarian secretions. To test this hypothesis, oestradiol-containing silicone elastomer capsules were implanted s.c. into juvenile rats, immediately after ovariectomy, and plasma prolactin profiles examined 2 days later (28 days of age).(ABSTRACT TRUNCATED AT 250 WORDS)     

Involvement of prolactin in the onset of puberty in female rats

Pimozide (1 mg/kg per day), bromocriptine (1 mg/kg per day) or domperidone (0.1 mg/kg per day) administered daily to rats from day 21 did not change the age at which vaginal opening occurred, nor did they affect the body weight at that age. Therefore the evolution of prolactin levels was different in these three groups. The pimozide-treated group showed high prolactin levels measured on day 23, at vaginal opening and at first oestrus. In the bromocriptine-treated group, levels were undetectable on the day of vaginal opening. Chronic treatment with domperidone failed to increase prolactin levels on day 23 and at vaginal opening. Nevertheless, large increases were observed after a single injection of domperidone at both 21 and 30 days of age. A significant increase in LH observed on day 23 in the pimozide-treated group was the only effect on gonadotrophin levels which was detected. Ovarian weights were unaffected by the treatments, whereas adrenal weight was increased in the bromocriptine-treated group and decreased in the pimozide- and domperidone-treated groups. Female rats grafted on day 21 with one additional pituitary gland from adult (90 days) or young (21 days) donors showed a similar advancement in the time of vaginal opening, although the animals bearing an adult pituitary gland showed higher prolactin levels than those observed in animals grafted with young pituitary glands. This study suggested that the onset of puberty is not closely linked with the evolution of prolactin levels and that the hormone itself is not indispensible for the process.     

Role of 17beta-estradiol and/or progesterone on insulin sensitivity in the rat: implications during pregnancy

The mechanism for the development of insulin resistance in normal pregnancy is complex and is associated with serum levels of both progesterone and 17beta-estradiol. However, it remains unclear whether estrogens alone or progestins alone can cause insulin resistance, or whether it is a combination of both which produces this effect. We attempted to determine the role played by progesterone and/or 17beta-estradiol on the phenomena of sensitivity to insulin action that take place during pregnancy in the rat. Ovariectomized rats were treated with different doses of progesterone and/or 17beta-estradiol in order to simulate the plasma levels in normal pregnant rats. A euglycemic/hyperinsulinemic clamp was used to measure insulin sensitivity. At days 6 and 11, vehicle (V)- and progesterone (P)-treated groups were more insulin resistant than 17beta-estradiol (E)- and 17beta-estradiol+progesterone (EP)-treated groups. Nevertheless, at day 16, the V, EP and E groups were more resistant to insulin action than the P group. On the other hand, the V, EP and E groups were more insulin resistant at day 16 than at day 6, whereas the P group was more insulin resistant at day 6 than at day 16. Our results seem to suggest that the absence of female steroid hormones gives rise to a decreased insulin sensitivity. The rise in insulin sensitivity during early pregnancy, when the plasma concentrations of 17beta-estradiol and progesterone are low, could be due to 17beta-estradiol. However, during late pregnancy when the plasma concentrations of 17beta-estradiol and progesterone are high, the role of 17beta-estradiol could be to antagonize the effect of progesterone, diminishing insulin sensitivity.