The roles of prolactin and testosterone in the development and function of granulosa lutein tissue in the rat

The luteotropic roles of prolactin and testosterone (or estradiol formed in luteal tissue) were investigated in hypophysectomized rats with homografts of granulosa lutein tissue. Using this approach, we could determine the effects of prolactin independently of estrogen, since granulosa lutein tissue does not produce estrogen de novo under these conditions. Luteinizing granulosa cells were expressed from the ovaries of immature pregnant mare's serum gonadotropin-primed Fischer 344 rats 6 h after injection of human chorionic gonadotropin. The cells were transplanted beneath the kidney capsule of adult, hypophysectomized, ovariectomized Fischer 344 recipients, which were treated with hormones daily for 12 or 14 days. In rats without treatment (no hormones, n = 3) and in rats treated with only testosterone (Silastic capsule, n = 6), only small amounts of luteal tissue (less than 5 mg/rat) were found and serum progesterone remained at low concentrations (10 ng or less) throughout the experiment. In contrast, in rats treated either with ovine prolactin (300 micrograms/day, n = 10) or with the combination of prolactin and testosterone (n = 12), serum progesterone increased to 43 ng/ml by Day 8. Beyond Day 8, serum progesterone continued to rise in rats treated with the combination of prolactin and testosterone to reach a mean value of 87 ng/ml by Day 14, and mean homograft wet weight was 49 mg/rat; in rats treated with only prolactin, serum progesterone decreased to 25 ng/ml by Day 14 and homograft wet weight was lower (24 mg/rat). Prolactin and testosterone together stimulated more homograft aromatase activity in vivo than did prolactin alone, but the in vitro production of progesterone was not different.(ABSTRACT TRUNCATED AT 250 WORDS)     

Implantation delay and anti-deciduogenic activity in the rat by the anti-androgen, hydroxyflutamide

Studies were undertaken to determine whether the anti-androgen, hydroxyflutamide, has anti-progestagenic activity by using implantation, maintenance of pregnancy, and decidualization as end points. Prepubertal rats were induced to ovulate with the injection of 4 IU pregnant mare's serum gonadotropin and allowed to mate. Mated females were assigned randomly to various treatment groups. Beginning at 0800 h on Day 4 of pregnancy and at 12-h intervals thereafter, rats received a series of 6 s.c. injections of 5 mg hydroxyflutamide in oil, or oil only. Localized changes in endometrial vascular permeability, indicative of implantation, were assessed on Days 6 and 8 of pregnancy, after an injection of Evans blue dye. By Day 6, implantation has been initiated in the vehicle-treated rats, but not in hydroxyflutamide-treated rats. Hydroxyflutamide treatment was terminated on Day 6, and implantation was initiated by Day 8. The weights of uterine dye sites in hydroxyflutamide-treated rats on Day 8 were similar to those in vehicle-treated rats on Day 6. The number of fetuses and placentae were similar in all groups on Day 19. The weights of fetuses in both hydroxyflutamide-treated and hydroxyflutamide + progesterone-treated rats were similar and significantly lower than those in control rats. Although there were no significant differences between vehicle-treated or hydroxyflutamide-treated rats in the proportion of rats delivering and litter size, the hydroxyflutamide-treated rats delivered pups a mean of one day later than did the controls. Endometrial decidualization in ovariectomized, steroid-treated rats, following artificial stimuli, was significantly suppressed in hydroxyflutamide-treated rats compared with controls.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effect of preoptic region implants of dilute estradiol on the maternal behavior of ovariectomized, nulliparous rats

Estrogen implanted directly into the medial preoptic region of pregnant Charles River Sprague-Dawley rats hysterectomized and ovariectomized on Day 16 of gestation mimics the effects of systemic estrogen treatment at this time by reducing the latency to respond to foster pups with maternal behavior (Numan, Rosenblatt, and Komisaruk, 1977). The present report describes the pup-directed responses of ovariectomized, nulliparous Zivic-Miller Sprague-Dawley rats that received bilateral medial preoptic implants of either cholesterol (n = 11) or estradiol diluted 1:10 with cholesterol (n = 11). Two days after treatment these animals were housed with three foster pups: their responsivity to pups and quality of nests built were then assessed, at first hourly and then daily. Rats receiving intracranial estradiol required significantly shorter exposures to pups than did cholesterol-treated animals before initiating carrying and grouping of 3 dispersed pups in a maternal nest during a 15-min test. On other measures, however, the groups did not differ (e.g., proportion grouping pups overnight, time required to complete retrieval of pups to the nest, time required to rebuild a disrupted nest). Animals treated with cholesterol and animals with estradiol implants did not differ in uterine weight at the time of sacrifice, suggesting that estrogen did not leak, even from this well-vascularized implant site, into the circulation. Thus, as in the pregnant animal, the facilitating effects of estrogen on maternal behavior can be mediated through the medial preoptic region; however, the effects were evident only when a test requiring retrieval of several pups within an arbitrarily short interval was given.     

Stimulation of maternal responsiveness after pregnancy termination in rats: Effect of time of onset of behavioral testing

To assess the effects of varying the time interval between surgical pregnancy termination and onset of behavioral testing on the expression of maternal behavior, both responsiveness to foster young and nest building-were measured in rats ovariectomized (O), hysterectomized (H), ovariectomized and hysterectomized (OH), or sham-operated (I) on Day 17 of pregnancy at 10:00 and first tested for maternal responsiveness 6, 24, or 48 hr after surgery. When behavioral testing was started 6 hr after surgery O and OH groups responded maternally to foster pups faster than H or I groups, and H females responded maternally faster than I animals (O = OH < H < I). In animals first tested 24 hr after surgery shorter latencies to retrieve and group foster young and nest build were found in the three pregnancy-terminated groups than in I animals, while the three pregnancy-terminated groups did not differ from one another with respect to either behavioral measure (O = OH = H < I). In contrast, when testing was initiated 48 hr after surgery, hysterectomized (H) animals responded maternally faster than did ovariectomized (O and OH) or intact pregnant (I) groups (H < OH < I, H < O = I). These data demonstrate that varying the time of onset of behavioral testing after surgical pregnancy termination affects elicitation of responsiveness to foster young and also affects nest building behavior in pregnancy-terminated animals. The differences in onset of maternal behaviors among pregnancy-terminated animals are discussed in relation to progesterone and estrogen secretion after surgery.     

Effect of estrogen and placental lactogen on lactogenesis in pregnant rats

The removal of the corpora lutea or ovariectomy on Day 18 of pregnancy induced a rise in serum prolactin 24 h after surgery with a rapid decline to control values 4 h after the surge, only in the ovariectomized group. When hysterectomy was performed in addition to luteectomy or ovariectomy a similar rise in prolactin was obtained. Lactose synthetase activity in mammary tissue was significantly higher in the luteectomized and luteectohysterectomized rats when compared with ovariectomized, ovariohysterectomized rats and the sham-operated group. Estrogen treatment 12 h after ovariectomy increased serum prolactin and lactose synthetase activity to values similar to those measured in luteectomized rats, but this increase was significantly greater when compared with the ovariectomized-nontreated group. Treatment with Tamoxifen did not decrease serum prolactin in the luteectomized rats but lactose synthetase was reduced to values similar to that obtained in ovariectomized rats. Treatment with 2 bromo-alpha-ergocryptine-mesylate (CB-154) prevented the rise in serum prolactin in the ovariectomized, luteectomized and luteectohysterectomized groups, but lactose synthetase activity was lowered to control values (sham-operated rats) only in the luteectohysterectomized rats. According to these findings, rat placental lactogen in the absence of prolactin and progesterone induces lactose synthesis. Estrogen facilitates prolactin but not placental lactogen action on lactose synthetase activity.     

Growth response, endocrine profiles and reproductive performance of fine-wool ewe lambs treated with ovine prolactin before breeding

Twenty-four 6-mo-old ewe lambs received one of two ovine prolactin (oPRL) treatments 28 d before fall breeding. Beginning on the first day of treatment (Day 0), 12 lambs received a subcutaneous injection (12 ml) of a carrier vehicle (0 mg oPRL) on alternate days for 28 d while 12 lambs received injections containing 5 mg oPRL. On Days 0 and 28, jugular blood was collected from six lambs in each group before treatment and at 30-min intervals for 6 h thereafter. Neither feed intake, efficiency of gain nor animal weights differed (P > 0.20) between groups. One hour after treatment on Day 0, ewe lambs receiving 5 mg oPRL had greater (P < 0.10) serum PRL levels than did controls (121.9 and 61.5 +/- 24.7 ng/ml, respectively). Differences in serum PRL persisted throughout remaining sampling intervals on both Days 0 and 28. Serum samples obtained on alternate days during the 28-d treatment period revealed no differences (P > 0.20) in PRL concentrations between control (48.3 +/- 5.3 ng/ml) and oPRL-treated (55.7 +/- 5.3 ng/ml) ewes. Neither serum insulin nor growth hormone responded (P > 0.05) to exogenous oPRL on either Day 0 or 28. No difference (P > 0.30) in percentage of ewe lambs cycling during treatment or breeding was detected between groups. Subsequent lambing percentages were similar (P > 0.30), with 36.4% of control and 25.0% of oPRL-treated ewes producing offspring. Administering 5 mg oPRL on alternate days for 28 d before breeding did not enhance growth and(or) reproductive performance in virgin ewe lambs.     

Dissociation between increased growth hormone and prolactin secretion during the morning hours of early pregnancy in the rat

We investigated whether serum growth hormone (GH) concentration changes in association with the rise in serum prolactin (PRL) concentration known to occur during the early morning hours in the pregnant rat. Animals were kept in a room with the lights on from 0500 to 1900 hours (hr) daily and decapitated for the collection of trunk blood at 2200 or 2400 hr on Day 6 of pregnancy or at 0200, 0400, 0800 or 1000 hr on Day 6 of pregnancy. Serum GH concentration rose more than 4-fold from low levels at 2200 and 2400 hr to higher levels at 0400 and 0800 hr and then declined by 1000 hr. Serum prolactin (PRL) concentration followed a similar pattern except that it returned to low levels earlier, by 0800 hr. Serum luteinizing hormone, follicle-stimulating hormone and thyroid-stimulating hormone concentrations showed no significant changes. Serum GH levels at 0800 hr in pregnant rats were higher than those observed in cyclic rats (13 time periods sampled). The results demonstrate that serum GH concentration is elevated during a circumscribed period in the 6- to 7-day pregnant rat. The time of onset of the rise is similar to that for serum PRL but the elevation in GH levels persists longer than that for PRL.     

Effects of estradiol and progesterone on early embryonic development in aging rats

Regularly cyclic, middle-aged female rats exhibit a decreased incidence of fertility, and those females that are fertile produce small litters. These decreases in fertility and litter size are associated with reduced numbers of normal blastocysts formed and implanted, suggesting that pre- and/or peri-implantation failures may be the causes for these aging-related reproductive declines. The present study examined the relationships and influence of circulating estradiol (E2) and progesterone (P) levels on early embryonic development and implantation in middle-aged rats. Serial blood samples obtained from cannulated, middle-aged pregnant rats revealed minor decreases in plasma P and increases in E2 levels during Days 2-4 of pregnancy, compared to young pregnant rats, resulting in significantly (p less than 0.001) decreased plasma P/E2 ratios. These alterations in endogenous hormone secretion in middle-aged pregnant rats were associated with fewer normal blastocysts on Day 5 of pregnancy and reduced numbers of normally implanting embryos. Correlation analysis further revealed a significant (p less than 0.05) inverse relationship between mean circulating E2 levels and numbers of normal conceptuses on Day 12 of gestation. Moreover, s.c. administration of P implants (in Silastic) to middle-aged pregnant rats increased serum P levels by about 34-40 ng/ml, and significantly (p less than 0.05) reduced the incidence of abnormal embryos before implantation. In contrast, treatment with E2 minipumps produced a sustained rise in serum E2 (by about 7-15 pg/ml) and resulted in the complete absence of embryos in the reproductive tracts by Day 5 of pregnancy.(ABSTRACT TRUNCATED AT 250 WORDS)     

Role of luteinizing hormone-releasing factor (LH-RF) and LH on maintenance of early gestation in rats.

The role of luteinizing hormone (LH) and LH-releasing hormone (LH-RH) in the maintenance of early pregnancy in rats was studied. Serum levels of progesterone (P) and LH were measured daily in untreated pregnant rats from Day 4 through parturition. Serum levels of P and LH were determined on Days 11 and 15 of pregnancy in animals treated with antisera to LH (LH-A/S) and to LH-RH (LH-RH-A/S) on Days 8-10. Serum levels of P peaked on Days 7 and 16 in untreated animals, after which they declined sharply just before delivery. Serum LH fluctuated between 30-160 ng/ml during pregnancy but did not exhibit any distinctive peaks. Treatment with .2 ml LH-A/S on Days 8-10 reduced serum P to virtually undetectable levels on Day 11, and only a slight recovery was evident on Day 15. Lower doses of LH-A/S had no effect. Administration of 1.3 ml LH-RH-A/S had no effect on serum levels of P or LH, and did not impede fetal development. The results indicate that LH is essential to the luteotropic complex of early pregnancy in the rat, and also that LH-RH-A/S can maintain to some extent basal levels of P and LH during early pregnancy.     

Critical progesterone requirement for maintenance of pregnancy in ovariectomized rats

The minimum progesterone concentration required to maintain the pregnancy was studied by varying doses of progesterone given subcutaneously to rats ovariectomized on Day 8 of pregnancy. Injecting 3 mg progesterone plus 200 ng oestradiol benzoate daily provided serum progesterone values between 25.4 +/- 7.0 and 35.2 +/- 6.2 ng/ml throughout Days 10-19 which were significantly lower than normal levels (P less than 0.05), but resulted in 93.6% of fetal survival on Day 19 which was not significantly different from 93.3% in the control group. Injecting 2 mg progesterone plus 200 ng oestradiol benzoate daily gave progesterone values between 13.2 +/- 4.6 and 19.0 +/- 6.2 ng/ml and could not maintain fetal viability to Day 19 (14.2%, P less than 0.05 compared with control group). Critical times to supplement progesterone in rats ovariectomized on Day 8 or Day 15 were studied by varying the time of progesterone implantation after ovariectomy. Progesterone implants were administered 8, 12 and 24 h after ovariectomy on Day 8 and 24, 36 and 48 h after ovariectomy on Day 15. On Day 8, progesterone replacement could be delayed to 8 h but not 12 h, while on Day 15, progesterone replacement could be delayed up to 36 h but not 48 h after ovariectomy without affecting fetal survival.     

Hormonal Responses of Male Gerbils to Stimuli from Their Mate and Pups

Following copulation and cohabitation with a pregnant female, male gerbils show high levels of parental behavior toward their pups. The initiation of male parental behavior may be the result of neuroendocrine changes induced by cohabiting with the pregnant female or by pup stimuli. Experiment 1 examines the changes in androgen and prolactin levels in male gerbils cohabiting with females over the reproductive cycle. Gerbils were mated and blood samples taken from males for hormone analysis 1, 10, and 20 days after pairing and 3, 10, and 20 days after pups were born. A group of unmated male gerbils served as controls. Plasma prolactin levels of males were elevated throughout the female's pregnancy and lactation periods, but were only statistically significantly higher than those of unmated males 20 days after pups were born. Androgen levels rose during pregnancy and dropped significantly after the birth of the pups. These hormonal changes are similar to those found in males of monogamous birds and differ from those found in males of polygynous rodents such as the rat. Experiment 2 examined the hormonal responses of male and female gerbils to pup replacement after 4 hr of parent–pup separation. Female gerbils showed a significant elevation of prolactin levels 1 hr after pup replacement, but males did not. Males with pups returned showed no difference in androgen levels from males who did not have pups returned. Thus, male gerbils show neuroendocrine changes following long-term cohabitation with their mate and pups, but do not show acute hormone responses to pup removal and replacement. These results indicate that parental males have neuroendocrine changes associated with parental behavior and these differ from the neuroendocrine changes underlying female parental behavior.     

Influence of progesterone concentrations on secretory functions of trophoblast and pituitary during the first trimester of pregnancy in dairy cattle

The essential role played by progesterone in the maintenance of pregnancy is unequivocal; however, the effects of progesterone on the secretory patterns of placental and pituitary molecules during the gestation period are not well defined. The objective of this study was to describe pregnancy-associated glycoprotein (PAG) concentrations (measured by RIA-497 and RIA-Pool) in pregnant females with progesterone concentrations lower (low-P4 group, n=20) or higher (high-P4 group, n=17) than the mean of 8.74 ng/mL on Day 21 (AI=Day 0). Luteinizing hormone (LH) and prolactin concentrations were also measured in both groups. Throughout the study period, blood samples were collected on Days 0, 21, 45, 60, and 80 from 37 females that were confirmed to be pregnant. PAG concentrations measured by both RIA-497 and RIA-Pool tended to be higher in high-P4 group than in low-P4 group from Day 30 until Day 80. On Day 80, plasma PAG concentrations that were measured using RIA-497 were observed to be higher (P<0.05) in the high-P4 group than in the low-P4 group (10.2+/-8.7 ng/mL versus 6.9+/-3.8 ng/mL). Concentrations of LH on Day 60 and prolactin on Day 80 were observed to be significantly lower (P<0.05) in the high-P4 group. There was a tendency for the concentrations of LH (Days 45 and 80) and prolactin (Days 30, 45, and 60) to be lower in cows in the high-P4 group than in the low-P4 group. Our results suggest the existence of a relationship among the concentration levels of progesterone, PAG, LH, and prolactin during early pregnancy.     

Nocturnal surges and reflexive release of prolactin in parentally behaving virgin female and male rats

Maternally behaving virgin rats are capable of releasing prolactin reflexively in response to stimulation by pups, especially during the proestrous/estrous phase of the cycle. When such rats are chronically exposed to pups they usually undergo a state of pseudopregnancy during which prolactin is secreted in a pattern of nocturnal surges. The present series of experiments evaluated the initiation of nocturnal prolactin surges in maternally behaving virgins, the role of estrogen in the reflexive release of prolactin, and the influence of gender on these two modes of prolactin secretion. It was found that the nocturnal surges of prolactin are already present on Days 1 and 2 of pup-induced pseudopregnancy. At this stage, however, the surges are not yet autonomous, seeing that pseudopregnancy is interrupted shortly after removal of the pups on Day 2. Activation by vaginocervical stimulation of the "mnemonic" neurogenic system that controls the autonomous nocturnal prolactin surges did not interfere with the reflexive pup-induced release of prolactin in maternally behaving virgins. The capacity of reflexive prolactin release in the virgin rat was abolished by ovariectomy, restored by estrogen replacement, and persisted for only 24 hr following estrogen removal. Paternally behaving males subjected to chronic exposure to pups were incapable of secreting nocturnal surges of prolactin characteristic of the pseudopregnant female. Such males were also incapable of releasing prolactin reflexively in response to stimulation by pups, even when supplemented with exogenous estrogen. These results indicate that the two modes of prolactin secretion are sex dependent, and that the maternally behaving virgin, unlike the postpartum rat, requires concurrent estrogenic facilitation for releasing prolactin in response to stimulation by young.     

Hormonal changes associated with estrogen induced luteolysis in the pregnant hamster: a reevaluation of the luteotropic complex

Hamsters were injected sc on Day 1 of pregnancy (sperm positive) with 50 micrograms estradiol cyclopentylpropionate (ECP) or peanut oil. On Day 5, serum progesterone (P4) was 10.6 ng/ml in controls vs 3.1 ng/ml after ECP. In the ECP group, serum prolactin (PRL) and follicle stimulating hormone (FSH) did not differ from controls but serum luteinizing hormone (LH) was significantly lower than that of the controls, and usually below the sensitivity of the radioimmunoassay (RIA). After ECP, structural signs of luteolysis (weight and histology) and absence of antral follicles characterized the ovary. Injection of an anti-LH serum on Day 4 halved serum P4 levels on Day 5 in control animals but caused no further lowering of P4 in ECP-treated hamsters. Treatment on Days 1-5 with 1.0 IU hCG or 10 micrograms LH plus ECP on Day 1 restored, by the afternoon of Day 5, serum P4 to the control range (9-10 ng/ml) and antral follicles were now present. The results indicate that a large dose of ECP causes luteolysis by reducing LH levels and reinforce the concept of a luteotropic complex in the hamster with PRL and FSH constituting the minimal components and LH serving as a synergist.     

Lactational [correction of Lacational] anovulation in rats and its dependency on progesterone

The relationship between prolactin (PRL) secretion and anovulation in lactating rats was studied. Normal lactating rats and lactating rats treated with antiserum against luteinizing hormone-releasing hormone at the time of postpartal ovulation were used. Normal lactating rats were treated with either a dopamine agonist (CB-154, 150 micrograms/rat) on Day 10 or 13, or pups removal on Day 7 or 10, and thereafter luteolysis and inhibition on PRL secretion were assessed. With the CB-154 treatment, the incidence of luteolysis increased as the lactational period advanced (42% vs 72%), whereas it decreased (73% vs 14%) with the pups removal. Thus, dopamine effectively inhibited PRL secretion during the later lactational stage, but could not do so during the earlier stage when there were mechanisms other than dopamine stimulating PRL secretion. Following luteal regression induced by CB-154, ovulation did not occur if the rats were treated with CB-154 on Day 10, whereas 50% of the rats ovulated within 4 days if treated on Day 13. Furthermore, in the lactating rats treated with anti-luteinizing hormone-releasing hormone serum during late pregnancy, ovulation was not observed until Day 10 of lactation. Since the serum progesterone levels were low in these rats due to the absence of ovulation and lactational corpora lutea, the blockade of ovulation was not due to elevated circulating progesterone during the early lactational period. The mechanism of ovulation blockade during lactation thus seems to shift from being progesterone independent to progesterone dependent at a similar period when the neuroendocrine control of PRL secretion shifts from dopamine independent to dependent.     

Radioimmunoassay of a bovine pregnancy-associated glycoprotein in serum: its application for pregnancy diagnosis.

A sensitive and specific double-antibody RIA for a bovine pregnancy-associated glycoprotein (bPAG) is described. The limit of detection was 0.2 ng/ml. The assay was specific for bPAG in that pituitary and placental gonadotropic hormones and other placental or serum proteins assayed in serial dilutions did not cross-react. The RIA allowed measurement of bPAG in placental extracts, fetal serum, fetal fluids, and serum or plasma of pregnant cows. About 20% of unbred heifers and nonpregnant cows had detectable levels ranging from 0.30 +/- 0.09 to 0.50 +/- 0.17 ng/ml (mean +/- SD), and 15% of bull sera showed higher concentrations (3.01 +/- 1.73 ng/ml) of bPAG or bPAG-like protein. Variations among animals was observed in fetal serum bPAG concentrations. Bovine PAG was detected in maternal peripheral blood at Day 22 of pregnancy (mean +/- SD, 0.38 +/- 0.13 ng/ml) in some animals and at Day 30 in all pregnant cows. Peripheral serum bPAG levels increased progressively to 3.60 +/- 1.73 ng/ml (mean +/- SD) at Day 30 of pregnancy, to 24.53 +/- 8.81 ng/ml at Day 120, and to 1551.91 +/- 589.68 ng/ml at Day 270. Peak concentration of bPAG was 2462.42 +/- 1017.88 ng/ml and it occurred 1-5 days prior to parturition. After delivery, bPAG concentrations decreased steadily to 499.63 +/- 267.20 ng/ml at Day 14 postpartum (pp), 10.12 +/- 7.84 ng/ml at Day 60 pp, and 1.44 +/- 1.08 ng/ml at Day 90 pp. The undetectable concentration (less than 0.20 ng/ml) was reached by Day 100 +/- 20 pp. An investigation undertaken in Holstein heifers, Holstein cows, and Hereford cows used as recipients for purebred Holstein embryos supplied evidence of the influence of breed of recipient and sex of fetuses on peripheral concentrations of bPAG. A herd of 430 Holstein-Friesian heifers that had received transferred embryos were bled at Day 35 postestrus (pe) for measurement of bPAG. The bPAG was detected in 287 of 430 serum samples analyzed. By rectal palpation performed at Day 45 pe, 267 heifers with detectable levels of bPAG at Day 35 pe were confirmed to be pregnant as were 3 of 143 heifers previously diagnosed as not pregnant by RIA. These results suggest that detection of this placental-specific antigen in the serum could be used as a specific serological method for early pregnancy diagnosis in cattle from 28 days after breeding.     

Relation of parity and estrous cyclicity to the biology of pregnancy in aging female rats

In the female rat, the incidence of regular estrous cyclicity and fertility decreases progressively during aging, and the causes for these are unknown. To reveal the biology of pregnancy in aging rats, we performed a longitudinal study in a colony of multiparous rats bred every 2 mo. Beginning at 4 mo and continuing to 12 mo of age in these same individual females, we determined the chronological changes in estrous cyclicity, examined the relationship between the estrous cycle pattern and fertility, and recorded the numbers of live and dead pups delivered at term. In separate groups of 4- to 12-mo-old multiparous rats, we counted the number of ova present in the oviducts (ovulation rate) one day after mating and the number of grossly normal blastocysts found in the uteri on Day 5 of pregnancy. Similar studies were also performed in primiparous rats of 8, 10, and 12 mo of age. The cessation of regular cyclicity during aging occurred significantly (p less than 0.01) earlier in virgin than multiparous rats. Fertility followed a similar but more dramatic pattern of decline than did the incidence of regular cyclicity in both the multiparous and virgin females. Few irregularly cyclic and persistent-estrous females had fertile gestations after mating, and increasing proportions of regularly cyclic females also failed to reproduce successfully at middle age (8-12 mo). Thus, regular ovulatory cycles were essential but not sufficient for fertile gestations in aging rats. Beginning at 6 mo of age, the litter sizes of multiparous rats decreased progressively, and these decreases were associated with a similar decline in the number of live but not dead pups delivered. Also, the percentage of dead pups/total number of pups delivered increased steadily during aging in multiparous (from 14% to 69%) but not primiparous females. The litter sizes of 8- to 10-mo-old primiparous females were not different from those of multiparous rats. However, the litter sizes of irregularly cyclic rats were consistently smaller than those of regularly cyclic females. Thus, parity had little effect on fecundity in aging females, whereas the cessation of regular ovulatory cycles during aging greatly decreased both the incidence of fertility and the litter size.(ABSTRACT TRUNCATED AT 400 WORDS)     

Progesterone inhibition of maternal behavior in the rat

The present series of experiments investigated the role of progesterone in inhibiting the onset of maternal behavior in the rat. Female rats hysterectomized and ovariectomized on Day 16 of pregnancy and injected subcutaneously with 20 μg/kg of estradiol benzoate (EB) show a short latency to onset of maternal behavior when presented with test pups 48 hr later. A subcutaneous injection of either 1 or 5 mg of progesterone on Day 16 of pregnancy and again 24 hr later inhibited this EB-induced short-latency onset of maternal behavior. The central neural site at which progesterone might act to produce this inhibitory effect was explored. Famale rats, hysterectomized and ovariectomized on Day 16 of pregnancy and injected subcutaneously with EB, received implants of crystalline progesterone on Day 16 of pregnancy into either the medial preoptic area, ventromedial hypothalamus, midbrain tegmentum, dorsal raphe nucleus, or median raphe nucleus. No inhibitory effects were found and all females showed a short-latency onset of maternal behavior. Several possible explanations for this lack of inhibitory effect of intracerebral implantation of progesterone are discussed.     

Suckling-induced prolactin release potentiates mifepristone-induced lactogenesis in pregnant rats

Suckling, starting at 19:00 h on Day 18 of pregnancy, induced a significant increase in serum prolactin concentration at 20:00 h on Day 19 of pregnancy, but no increase in mammary gland casein or lactose content. Mifepristone (2 mg/kg) injection at 08:00 h on Day 19 of pregnancy induced significant increases in casein, but not in lactose, 24 h after administration. Mifepristone alone did not induce prolactin secretion, indicating that lactogenesis was induced by placental lactogen in the absence of progesterone action. When mifepristone was injected into suckling rats, serum prolactin concentrations were higher than in the untreated suckling rats. Casein in these rats increased significantly 12 h after mifepristone administration and lactose at 24 h after. If the suckling mifepristone-treated rats were given two injections of bromocriptine (1.5 mg/kg) at 12:00 h on Days 18 and 19 of pregnancy, serum prolactin concentrations were not increased by suckling, but casein and lactose concentrations in the mammary gland showed values similar to those obtained in the mifepristone-treated non-suckling rats. Mifepristone can therefore potentiate suckling-induced prolactin release in pregnant rats, demonstrating a direct central inhibitory action of progesterone on prolactin secretion. This suckling-induced prolactin secretion, unable to induce casein or lactose synthesis in the presence of progesterone, enhanced significantly synthesis of these milk components in the absence of progesterone action (rats treated with mifepristone). Fatty acid synthase, which is stimulated by the suckling stimulus in lactating rats, was not modified by mifepristone or suckling in pregnant rats.     

Infertility in a line of mice with the high growth mutation is due to luteal insufficiency resulting from disruption at the hypothalamic-pituitary axis.

Animals with extreme body growth frequently experience poor reproductive performance, but the cause for this association has not been clearly established. A line of mice homozygous for the high growth (hg) mutation, a mutation that has a major effect on post-weaning growth, exhibits several reproductive deficits including an abnormally high incidence of luteal failure. The objective of the present study was to investigate luteal failure in high-growth (HG) mice during pregnancy and to determine whether the cause of the apparent luteal failure resides in the ovary or the hypothalamic-pituitary unit. In HG females with a demonstrated history of infertility, progesterone injections (1 mg s.c. daily) beginning on Day 1 postcoitum (p.c.) increased the proportion of animals pregnant at Day 17 of gestation. Twice-daily injections of 100 microgram of ovine prolactin (PRL) in alkaline saline given to another group of females beginning on Day 1 p.c. increased the proportion of HG females that were pregnant on Day 6 of gestation compared with saline-injected HG females, although PRL did not increase the pregnancy rate in HG females when compared with a group of noninjected control females. When ovaries from HG females were transplanted into ovariectomized congenic C57 hosts, the C57 graft hosts displayed normal estrous cycles, and upon mating the majority of graft hosts became pregnant. In contrast, when ovaries from C57 females were transplanted into ovariectomized HG hosts, the HG graft hosts displayed normal estrous cycles, but upon mating most were unable to maintain pregnancy. These results suggest that the hypothalamic-pituitary unit of the HG female provides an inadequate signal to the ovaries to maintain pregnancy. Luteal failure in HG females may be due to insufficient PRL as required for establishment and early maintenance of the CL during pregnancy in mice.