Regulation of the proestrus surge of prolactin secretion by gonadotropin and estrogens in the rat.
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Systemic levels of estradiol (E2) during the 4-day estrous cycle rise in a biphasic manner; slow elevations during diestrus II are followed by rapid increases during the early morning of proestrus (Kalra and Kalra, 1973). The relation of these alterations in systemic E2 levels to the proestrus surge of LH was investigated. Administration of progesterone (P 4 mg/rat) to rats at 2200 hr of diestrus II blocked ovulation and the surge of LH in the afternoon of proestrus. Injection of estradiol benzoate (EB, 5 mug/rat) 2 hr prior to, together with or following P treatment at 0300 hr of proestrus restored ovulation and the surge of LH. However, blockade of ovulation was not reversed if EB was administered at 0500 or 0800 hr of proestrus following P treatment on diestrus II. Similarly, bilateral ovariectomy at 2300 hr of diestrus II but not at 0300 hr or later on proestrus abolished the proestrus surge of LH. Apparently, the rapid rise in systemic E2 between 2300 hr of diestrus II and 0300 hrs of proestrus is crucial and serves to activate the neural "trigger" of preovulatory release of LH. The role of ovarian E2 secretion prior to 2300 hr of diestrus II in facilitation of neural "trigger" was studied. In the absence of endogenous E2 priming produced by ovariectomy at 0800 hr of diestrus II, EB injections at 2300 hr of the same day failed to restore the proestrus surge of LH. However, when ovariectomy was delayed to allow normal E2 priming through diestrus II, administration of EB shortly after ovariectomy at 2300 hr of diestrus II elicited the normal proestrus surge of LH. These results indicated that following the initial "priming" of central site(s) with low levels of circulating E2 during diestrus II, rapid elevations in ovarian estrogen secretion between 2300 hr of diestrus II and 0300 hr of proestrus facilitated the neural "trigger" of pituitary LH release during the critical period on proestrus.
Changes in hypothalamic concentration and turnover index (TI) of norepinephrine (NE), dopamine (DA), and serotonin (5HT) were studied during the first estrous cycle at the onset of puberty. Rats were killed when 40% of the population showed open vaginas. They were classified according to the state of their reproductive tract and serum LH values, as in anestrus, early proestrus, late proestrus, estrus, or diestrus. Serum LH, PRL, and hypothalamic LHRH were measured by RIA. The TIs for DA and NE were measured by estimating their rate of decrease after administration of alpha-methyl-para-tyrosine, and for 5HT-TI by its increase after pargyline administration. An increase in NE-TI in the hypothalamus was observed between anestrus and early proestrus, a decrease in DA-TI during early and late proestrus, and a small increase in 5HT-TI between early and late proestrus. LH and PRL peaks were observed at late proestrus when DA-TI and NE-TI reached their lowest values and hypothalamic LHRH concentration was failing. After late proestrus, the DA-TI rose sharply, NE-TI remained essentially unchanged. 5HT-TI fell, and serum LH and PRL declined to basal levels. These results suggest that the LH and PRL surges at late proestrus during the first estrous cycle at puberty are associated with an increase in NE-TI at early proestrus, a progressive decline in DA-TI at early and late proestrus, and an increase in 5HT-TI at late proestrus.
Fluctuations of ovarian biosynthetic activity and effects of exogenous LH and FSH on it during the estrous cycle were investigated by measuring in vitro transfer rates of 14C from 14C-1-acetate into progesterone (P), 20 alpha-hydroxy-pregn-4-en-3-one (20 alpha-OH-P) and estrogen (estradiol and estrone, E) in the ovarian homogenates from rats autopsied at 2 hour intervals. The transfer rate of 14C from 14C-1-acetate into P was lowest in the afternoon of estrus and increased from the morning of diestrus 1, making its peaks during the afternoon of diestrus 2 and in the midnight of proestrus. The transfer of 14C into 20 alpha-OH-P was high on the days of diestrus 2 and proestrus with its peak in the afternoon of the latter day. The maximum transfer of 14C into E in the afternoon of proestrus and a high rate in the morning of estrus with relatively low one in the midnight were observed. Exogenously injected LH (150 mug) or FSH (150 mug) was either stimulatory or inhiibitory to the transfer rates of 14C from 14C-1-acetate into ovarian steroids. During day time of diestrus 2 and midnight between proestrus and estrus, the transfer of 14C into P and 20 alpha-OH-P increased by LH, and during day time of proestrus and from the afternoon of estrus to the morning of diestrus 1 decreased. The transfer of 14C into E increased by LH from the afternoon of diestrus 2 to the morning of proestrus, and decreased during the afternoon of proestrus and from the afternoon of estrus to the morning of diestrus 2. Administration of FSH was also stimulatory or inhibitory. The 14C transfer into P and 20 alpha-OH-P increased by FSH from the afternoon of estrus to the morning of proestrus, but in the afternoon of proestrus they decreased. Transfer of 14C into E increased by FSH significantly on the days of diestrus 2 and proestrus, and slightly on the day of estrus, while it decreased in the afternoon of diestrus 1.
We sought to determine whether the estrous phase of FSH release in cyclic female rats is dependent on the immediate presence of the diencephalon. A piece of forebrain, the diencephalon and part of the telencephalon, was surgically removed from female rats between 1130--1300 or 2000-2200 h on proestrus. Blood was withdrawn through indwelling venous cannulae during the afternoon and evening of proestrus and the early morning of estrus for RIA of plasma LH, FSH, and PRL concentrations. In rats sham operated at either time period, plasma LH, FSH, and PRL levels rose from 1345 to 1800 h on proestrus. Whereas the plasma LH and PRL concentrations fell from 1800 h on proestrus to 0300 h on estrus, the plasma FSH concentrations remained elevated during this period. The removal of the piece of forebrain around noon on proestrus blocked the rises in the plasma LH and FSH levels and caused high plasma PRL concentrations from 1345 h on proestrus to 0300 h on estrus. The removal of the piece of forebrain during the evening of proestrus did not interfere with the fall in plasma LH concentrations or the maintenance of elevated plasma FSH concentrations during either late proestrus or the early morning estrus, but did cause high plasma PRL levels during that time interval. Hypophysectomy combined with removal of the forebrain piece during the evening of proestrus resulted in a drop in plasma FSH and PRL concentrations. The results confirm that in the rat, 1) the prosencephalon plays an acute stimulatory role n causing the preovulatory LH surge and the proestrous phase of FSH release, 2) the prosencephalon exerts effects during the afternoon and/or early evening of proestrus that cause the estrous phase of FSH release, and 3) the estrous phase of FSH release occurs in the absence of acute diencephalic stimulation.
The roles of recombinant human inhibin A (rh-inhibin A) and rh-activin A in regulating the pituitary and ovary of the adult female rat were examined. Serum and pituitary FSH and LH and serum estradiol and progesterone concentrations were evaluated at 1, 2, and 6-12 h after sc hormone administration on metestrus and on proestrus. A second study examined the effect of the hormones 24 h after injection at 1000 h on each day of the cycle. Rh-inhibin A inhibited FSH secretion 60 min after injection on proestrus but did not alter serum FSH concentration on metestrus. FSH remained low for the 12 h examined during the evening of proestrus and on the morning of estrus. Serum LH concentration, pituitary FSH content, and pituitary LH content were not significantly changed under any experimental condition. Rh-inhibin A did not regulate estradiol concentration on metestrus or on proestrus; however, it did cause a rise in serum estradiol in animals treated on metestrus and diestrus and examined 24 h later. This suggests that inhibin may participate in regulating follicular maturation in a subset of developing follicles. Last, after rh-inhibin A treatment, the duration of the proestrus progesterone surge was shortened. Serum FSH concentration rose by 6 h after rh-activin A administration on proestrus and both FSH and LH rose by 6 h after hormone administration on metestrus. Rh-activin A significantly increased serum estradiol through 6 h of treatment on proestrus. Progesterone levels were significantly greater in animals treated on metestrus and killed 24 h later. The increased length of the midcycle progesterone surge may be the result of increased LH on metestrus. These studies suggest that rh-inhibin A and rh-activin A may regulate ovarian and pituitary function in a cycle dependent manner. Specifically, rh-inhibin A can acutely regulate FSH and progesterone on proestrus and estradiol during follicular development. Rh-activin A acutely regulates FSH on both proestrus and metestrus and LH on metestrus. Whether circulating endogenous inhibin or activin participate physiologically in these functions is under investigation.
Brainstem catecholaminergic neurons are believed to play an important role in the activation of luteinising hormone-releasing hormone (LHRH) neurons on the afternoon of proestrus which results in the luteinising hormone (LH) surge. To examine the respective roles of brainstem A1 and A2 neurons and the adjoining C1 and C2 adrenergic cells at this time, we have examined the patterns of Fos-immunoreactivity within tyrosine hydroxylase (TH) and phenylethanolamine-N-methyl transferase (PNMT) neurons during diestrus and proestrus. Initial studies demonstrated that the LH surge commenced at approximately 15:00 h in proestrous animals and that peak plasma levels of LH were observed between 16:00 and 17:00 h. Groups of cycling female rats (n = 6) were then perfused between 09:00 and 11:00 (diestrus early) and 18:00 to 19:30 h (diestrus late) on diestrus and at the same times on proestrus (proestrus early and proestrus late). Double-labelling immunocytochemistry revealed little Fos expression by adrenergic neurons of the C1 or C2 cell groups and this did not change significantly between any of the experimental groups. Analysis of the A2 region was divided into rostral, middle and caudal divisions and all regions showed a significant (P < 0.01) increase in the number of Fos-expressing TH neurons (up to 35% of TH cells) in proestrus early animals compared with diestrus and proestrus late rats. In the A1 region, a significant increase in the number of TH neurons expressing Fos (approximately 33%) was detected in both proestrus early (P < 0.05) and diestrus early (P < 0.01) rats compared with animals perfused in the late afternoon (approximately 12%).(ABSTRACT TRUNCATED AT 250 WORDS)
The effect of mifepristone, a potent progesterone receptor antagonist, on the reproductive function during early pregnancy in rats was examined. A single dose of this drug (10 mg/kg) was injected s.c. at 1200 h on day 4 (Group 1), day 7 (Group II) or day 10 (Group III) of pregnancy. Gestation was interrupted and the vaginal cytology showed a typical proestrus condition two days after mifepristone treatment in all groups. When compared with cycling proestrus rats, serum LH concentrations at 1800 h in the mifepristone-induced proestrus were lower in Group I, similar in Group II and higher in Group III, and serum prolactin (PRL) values were lower in Group I, but not different in Groups II and III. Serum progesterone levels were higher in the three experimental groups when compared with cycling proestrus rats, and similar to that of pregnant rats. Rats in Group I showed a significantly lower sexual receptivity and ovulation rate when compared with Groups II and III or cycling proestrus rats. Most of the mifepristone-treated rats that copulated during the night of the induced proestrus did not become pregnant and showed a delayed pseudopregnancy-like condition. These results indicate that mifepristone administered in a single dose to early pregnant rats terminates pregnancy and induces a proestrus condition two days after treatment followed by successful postovulatory contraception. The mifepristone-induced proestrus is characterized by a differential pattern of serum LH, PRL and progesterone concentrations, mating behavior and ovulation rates, depending on the day of pregnancy when mifepristone is administered.
Ovaries isolated from rats in different stages of the sex cycle were explored for spontaneous or drug-induced contractile activity. The number of spontaneously active ovaries as well as the magnitude of the isometrically developed tension and frequency of contractions were greater during the periovulatory interval (late proestrus and estrus) than during early proestrus or metestrus. Furthermore, during estrus or late proestrus the left ovaries exhibited a mechanical activity significantly greater than that of the right ovaries. The oxytocin-triggered motility was clearly more marked immediately prior to ovulation (late proestrus) and greater in left ovaries than in right ovaries. In contrast, the contractions induced by prostaglandin F2alpha were similar during early proestrus and late proestrus. Ovarian contractile reactivity to norepinephrine indicated the presence in the tissue of alpha- and beta-adrenergic receptors. During early proestrus this agent stimulated the motility of left and right ovaries, whereas close to the ovulatory interval (late proestrus) it depressed the contractions of left ovaries. This last influence was blocked by propranolol. The existence of a close relationship between ovarian contractile activity and ovulation is reinforced by the present results in the rat. A tentative participation of oxytocin is also suggested. In addition, the influences of other possible regulatory agents of ovarian contraction, such as catecholamines and prostaglandins, are presented and discussed.