Persistent Estrous Research Articles

Ovarian Steroidogenic Responsiveness to Exogenous Gonadotropin Stimulation in Young and Middle-Aged Female Rats

Reproductive aging in the female rat is associated with gradual declines in LH secretion and ovarian progesterone (P) production. This study examined whether the influences of aging on P levels reflect decreased ovarian responsiveness to gonadotropin stimulation, as opposed to changes in gonadotropin release. Young and middle-aged regularly cyclic female rats received sodium pentobarbital to block endogenous proestrous luteinizing hormone (LH) surges, followed by administration of various doses of human chorionic gonadotropin (hCG). Similar treatments were performed in middle-aged acyclic persistent-estrous (PE) females. Injection of hCG resulted in equivalent plasma hCG levels in each treatment group. At the lowest hCG dose tested, a significant rise in plasma P levels was observed in middle-aged cyclic rats, but not in young cyclic or middle-aged PE females. This unexpected finding may reflect accelerated follicular development in middle-aged cyclic females, as suggested by a previous study. At the inte...

Changes of Cyclic AMP Levels and Phosphodiesterase Activities in the Rat Ovary

Cyclic AMP (cAMP) is a second messenger that plays a critical role in follicular recruitment, development and luteinization in the mammalian ovary. The cellular level of cAMP is largely dependent on the activity of phosphodiesterase (PDE), which degrades cAMP into 5'-AMP. The present study was conducted to investigate the level of cAMP and the activity of cAMP-PDE in postnatal rats; immature rats during gonadotropin-primed follicular development, ovulation and luteinization; adult rats during normal estrous cycling; and aged rats that spontaneously developed persistent estrous (PE) by radioimmunoassay (RIA). All four rat models were confirmed by histological examination of one ovary and assayed using the other ovary by RIA. In the postnatal rats, the ovarian cAMP level was high on day 10 after birth, while ovarian cAMP-PDE activity was highest at 21 days of age. In the immature female rats, both the ovarian cAMP level and cAMP-PDE activity increased remarkably after treatment with equine chorionic gonadotropin (eCG), increased continuously 24 h after injection of human chorionic gonadotropin (hCG) for induction of ovulation and luteinization, and then declined significantly. In the adult rats during the normal estrous cycle, the ovarian cAMP levels were low on the day of estrus, and there were no significant changes in ovarian cAMP-PDE activity throughout the estrous cycle. In the PE rats, the ovarian cAMP levels were similar to those of the adult rats on the day of estrus but were lower than those on the other days of the estrous cycle; ovarian cAMP-PDE activity was lower than that in the adult rats on any day of the estrous cycle. Together, these findings indicate that the ovarian cAMP level and cAMP-PDE activity were regulated in a stage-dependent manner during ovarian follicular development, atresia and luteinization and providing evidences that cAMP and cAMP-specific PDEs are involved in these physiological processes.

Temporal changes occur in the neuroendocrine control of gonadotropin secretion in aging female rats: role of progesterone.

The present study examined the gonadotropin surge-inducing actions of estradiol (E(2)), both alone and with progesterone (P(4)), in middle-aged, early persistent-estrous (PE) female rats that had become PE within 35 days. In addition, we also assessed the effect of P(4) on the mating-induced gonadotropin surges in these acyclic animals. Early PE rats were ovariectomized and received E(2) implants (Day 0). On Day 4, an s.c. injection of P(4) (0.5 mg/ 100 g body weight) at 1200 h markedly increased plasma P(4) and elicited both LH and FSH surges, whereas vehicle-treated controls displayed no rise in P(4) or gonadotropins. This observation confirms that at middle age, female rats no longer respond to the positive-feedback stimulation of E(2) on gonadotropin surges whenever the estrous cyclicity ceases. As PE continued, such a surge-inducing action of E(2) plus P(4) became diminished after 75 days of PE and disappeared thereafter. When caged with males, vehicle-treated early PE rats display a mating-induced increase in P(4) from the adrenal along with small gonadotropin surges. The amplitude of these mating-induced gonadotropin surges was enhanced by supplementation with exogenous P(4) in early PE rats. Our findings indicate that during the early phase of PE, the surge-inducing action of E(2) and P(4) remains intact but deteriorates as PE continues. Thus, a deficiency in P(4) secretion during aging may contribute to the diminished gonadotropin surge response in the hypothalamic-pituitary axis and the subsequent cessation of estrous cyclicity.

Chronic elevation of estradiol in young ovariectomized rats causes aging-like loss of steroid-induced luteinizing hormone surges.

This study was designed to test the hypothesis that the loss of LH surges in response to the stimulatory actions of estradiol and progesterone in middle-aged, persistent-estrous (PE) rats may be caused by chronic elevations in circulating estradiol. Five groups of regularly cycling young rats received an s.c. estradiol implant immediately after ovariectomy (Day 0). For determination of LH surges, blood samples were collected hourly between 1200-1900 h from each of the five groups at one of the following times: 3 days, or 1, 2, 4, or 8 wk later. On the next day, either progesterone (0.5 mg/100 g BW) or corn oil was injected s.c. at 1200 h, and samples were obtained as before. Incidence and amplitude of estradiol-induced LH surges decreased during the first 2 wk of estradiol treatment, after which no surges occurred. Progesterone enhanced the incidence and amplitude of estradiol-induced LH surges thus delaying their disappearance. These results support our hypothesis and demonstrate that the stimulatory actions of estradiol and progesterone on the LH surge sequentially diminish with time after exposure to estradiol in young rats. Thus, young rats chronically treated with estradiol may be a useful model for studying the mechanisms whereby LH surges are abolished in middle age during the hyperestrogenic state of PE.

Ovarian steroidogenic responsiveness to exogenous gonadotropin stimulation in young and middle-aged female rats.

Reproductive aging in the female rat is associated with gradual declines in LH secretion and ovarian progesterone (P) production. This study examined whether the influences of aging on P levels reflect decreased ovarian responsiveness to gonadotropin stimulation, as opposed to changes in gonadotropin release. Young and middle-aged regularly cyclic female rats received sodium pentobarbital to block endogenous proestrous luteinizing hormone (LH) surges, followed by administration of various doses of human chorionic gonadotropin (hCG). Similar treatments were performed in middle-aged acyclic persistent-estrous (PE) females. Injection of hCG resulted in equivalent plasma hCG levels in each treatment group. At the lowest hCG dose tested, a significant rise in plasma P levels was observed in middle-aged cyclic rats, but not in young cyclic or middle-aged PE females. This unexpected finding may reflect accelerated follicular development in middle-aged cyclic females, as suggested by a previous study. At the intermediate dose, young and middle-aged cyclic but not PE rats displayed significantly increased P in response to hCG. At the highest dose tested, all three groups of rats displayed increased P levels after hCG stimulation. However, P concentrations were significantly lower in middle-aged PE than regularly cyclic females. Northern and slot blot hybridization analyses revealed that ovarian mRNA levels for cytochrome P450 side-chain cleavage, the rate-limiting enzyme in P synthesis, were markedly reduced in PE rats following hCG stimulation. These findings indicate that ovarian responsiveness to gonadotropin stimulation is impaired in middle-aged PE, but not regularly cyclic rats, and suggest influences of cycle status on the biochemical and molecular mechanisms regulating ovarian steroid production. Furthermore, these findings reveal that attenuated P production in middle-aged proestrous rats is due to attenuated preovulatory LH surges, rather than decreased ovarian sensitivity to LH.

Altered regulation of pituitary luteinizing hormone secretion by GnRH and inhibin in the aged persistent-estrous female rat

The physiologic regulation of pituitary LH secretion is under the direct control of hypothalamic GnRH as well as feedback control via ovarian steroid and peptide hormones. Changes within the hypothalamic-pituitary-ovarian axis during reproductive aging lead to the gradual attenuation and the eventual loss of cyclic LH release that is seen in the old persistent-estrous rat. In the following studies, we determined whether changes in the pituitary response to regulation by hypothalamic GnRH and/or ovarian inhibin could be involved in age-related deficits in LH release by examining the direct effect(s) of GnRH and inhibin on LH release in old persistent-estrous (PE) and young female rats using an in vitro pituitary cell culture system. Both pituitary LH content and LH release were significantly lower in PE pituitary cells compared to young pituitary cells. GnRH (10 ng/ml) significantly increased LH release above basal levels in young and PE pituitaries. However, the magnitude of LH release was significantly lower in PE pituitaries compared to young pituitaries. Treatment with recombinant inhibin (0.1, 1, or 5 ng/ml) did not significantly affect basal or GnRH-stimulated LH release from young rat pituitaries under our culture conditions. In contrast, there were significant decreases in the levels of both basal LH and GnRH-stimulated LH release from PE pituitaries after treatment with 1 or 5 ng/ml human recombinant inhibin, as well as with inhibin plus 10 ng/ml GnRH, respectively. These results suggest that there are significant changes in the pituitary response to GnRH and ovarian inhibin regulation that may lead to significant alterations in LH secretion during aging.

Alterations in plasma and ovarian immunoreactive inhibin during reproductive aging in the female rat

Previously, we showed that ovarian inhibin α- and β;(A)-subunit mRNAs are elevated in middle-aged and old persistent-estrous (PE) female rats. To determine whether higher inhibin subunit mRNA expressions result in increased circulating inhibins during reproductive aging, plasma immunoreactive inhibin α (ir-inh α) and gonadotropins were measured in young, middle-aged and PE rats. Plasma LH profiles were distinctly divergent in the middle-aged rats with some showing LH surges indistinguishable from young rats and others showing significantly attenuated LH surges. Plasma ir-inh α in middle-aged rats with LH surges were similar to those of young rats. However, animals with attenuated LH surges had higher peak ir-inh α levels than young and middle-aged animals with LH surges. Immunohistochemistry revealed increased levels of ovarian inhibin α staining in those animals with attenuated LH surges. Overall, the highest plasma and ovarian inhibin α were found in PE rats which lack LH surges. However, significant decreases of plasma and ovarian inhibin α were seen after reinstatement of estrous cyclicity with progesterone implant treatment. Thus, increases in both plasma and ovarian inhibin α appear to be closely associated with attenuation or loss of the preovulatory gonadotropin surge that occurs during aging.

Apparent absence of negative feedback in middle-aged persistent-estrous rats following luteinizing hormone-releasing hormone agonist treatment: relation to plasma inhibin and 17 beta-estradiol.

Reproductive aging in female rats is associated with a transition from regular estrous cyclicity to an anovulatory condition described as persistent estrous (PE). This PE condition is characterized by continued follicular development with elevated circulating levels of estrogen and FSH. In an attempt to investigate further the age-related changes in neuroendocrine function of PE rats, we have developed a model through which the return of hypothalamic-pituitary and ovarian function can be assessed following the withdrawal of chronic LHRH agonist suppression. Subsequent to withdrawal of continuous (2.5 micrograms/h for 12 days) LHRH agonist [DTrp6, Pro9-NHEt]-LHRH (LHRH-AG) treatment, circulating FSH concentrations in PE rats increase and remain elevated with an apparent absence of ovarian negative feedback, and these rats fail to return to estrous cyclicity. In the present studies, estrogen administration induced significant decreases in FSH secretion in PE rats following withdrawal of LHRH-AG treatment and ovariectomy (OVX), suggesting that the negative feedback response to estrogen is maintained in PE females. However, progesterone administration 2 days later failed to elicit a positive feedback response of gonadotropin secretion in PE females prior to LHRH-AG treatment, serum inhibin and 17 beta-estradiol (E2) concentrations were similar in middle-aged PE rats and young cyclic females on proestrus, while FSH levels were significantly greater in PE rats. After withdrawal of LHRH-AG treatment, plasma FSH concentrations remained elevated in PE rats as compared to young rats despite similar increases in E2. However, increases in plasma inhibin were delayed and significantly attenuated in PE rats.(ABSTRACT TRUNCATED AT 250 WORDS)

Inhibin subunit gene expression and distribution in the ovaries of immature, young adult, middle-aged, and old female rats.

In young adult female rats, the patterns of inhibin subunit mRNA expression during the estrous cycle are regulated by cyclic changes in gonadotropin secretion and follicular development. Since there are distinct alterations in profiles of both hormone secretion and folliculogenesis during the reproductive lifespan of the female rat, we have characterized the gene expression and distribution of inhibin subunit mRNAs in immature (22 days), young adult cyclic (3-4 months), middle-aged cyclic (9-10 months), and old (12-13 months) persistent estrous (PE) rat ovaries. Northern blot analyses revealed that in contrast to young adult cyclic rats, inhibin alpha- and beta A-subunit mRNA levels in the ovaries of middle-aged cyclic rats remained substantially elevated after the proestrous gonadotropin surges and ovulation. Likewise, acyclic immature and old PE rats showed high levels of inhibin alpha- and beta A-subunit transcripts in their ovaries. In situ hybridization analyses demonstrated that inhibin alpha- and beta A mRNAs were abundantly expressed in maturing follicles of both young and middle-aged cyclic females, while high levels of alpha-subunit transcripts were only detected in the ovarian stroma of middle-aged animals. The ovaries of old PE rats had numerous large cystic follicles (with variable layers of granulosa cells) and few degenerating cyst-like structures (completely devoid of granulosa cells). Inhibin subunit transcripts were expressed abundantly in both the granulosa (alpha and beta A-subunits) and theca interna (alpha-subunit only) layers of large follicles, but were absent from degenerating cysts devoid of granulosa cells. The ovarian stroma of PE rats also expressed very high levels of inhibin-alpha, but not beta A mRNA. The ovaries of immature rats contained large numbers of uniformly developing secondary follicles. High levels of inhibin-alpha mRNA were expressed homogeneously in the granulosa layer of all growing follicles, whereas inhibin beta A mRNAs were only detected in selectively larger follicles with multiple layers of granulosa cells. Hormone RIAs of serum samples from these same groups of animals showed that basal levels of serum FSH were substantially higher in immature, middle-aged cyclic, and old PE rats than in young adult rats. These results demonstrate that enhanced ovarian inhibin subunit gene expression in the female rat is associated with increased serum FSH levels regardless of chronological age. On the other hand, aging appears to selectively enhance inhibin alpha, but not beta A, gene expression in the ovarian stroma, such that it may gradually become a major secondary site of alpha-subunit mRNA production in addition to the follicular compartments.

Inhibin subunit gene expression and distribution in the ovaries of immature, young adult, middle-aged, and old female rats

In young adult female rats, the patterns of inhibin subunit mRNA expression during the estrous cycle are regulated by cyclic changes in gonadotropin secretion and follicular development. Since there are distinct alterations in profiles of both hormone secretion and folliculogenesis during the reproductive lifespan of the female rat, we have characterized the gene expression and distribution of inhibin subunit mRNAs in immature (22 days), young adult cyclic (3-4 months), middle-aged cyclic (9-10 months), and old (12-13 months) persistent estrous (PE) rat ovaries. Northern blot analyses revealed that in contrast to young adult cyclic rats, inhibin alpha- and beta A-subunit mRNA levels in the ovaries of middle-aged cyclic rats remained substantially elevated after the proestrous gonadotropin surges and ovulation. Likewise, acyclic immature and old PE rats showed high levels of inhibin alpha- and beta A-subunit transcripts in their ovaries. In situ hybridization analyses demonstrated that inhibin alpha- and beta A mRNAs were abundantly expressed in maturing follicles of both young and middle-aged cyclic females, while high levels of alpha-subunit transcripts were only detected in the ovarian stroma of middle-aged animals. The ovaries of old PE rats had numerous large cystic follicles (with variable layers of granulosa cells) and few degenerating cyst-like structures (completely devoid of granulosa cells). Inhibin subunit transcripts were expressed abundantly in both the granulosa (alpha and beta A-subunits) and theca interna (alpha-subunit only) layers of large follicles, but were absent from degenerating cysts devoid of granulosa cells. The ovarian stroma of PE rats also expressed very high levels of inhibin-alpha, but not beta A mRNA. The ovaries of immature rats contained large numbers of uniformly developing secondary follicles. High levels of inhibin-alpha mRNA were expressed homogeneously in the granulosa layer of all growing follicles, whereas inhibin beta A mRNAs were only detected in selectively larger follicles with multiple layers of granulosa cells. Hormone RIAs of serum samples from these same groups of animals showed that basal levels of serum FSH were substantially higher in immature, middle-aged cyclic, and old PE rats than in young adult rats. These results demonstrate that enhanced ovarian inhibin subunit gene expression in the female rat is associated with increased serum FSH levels regardless of chronological age. On the other hand, aging appears to selectively enhance inhibin alpha, but not beta A, gene expression in the ovarian stroma, such that it may gradually become a major secondary site of alpha-subunit mRNA production in addition to the follicular compartments.

Pro-Gonadotropin-Releasing Hormone (ProGnRH) and GnRH Content in the Preoptic Area and the Basal Hypothalamus of Anterior Medial Preoptic Nucleus/Suprachiasmatic Nucleus-Lesioned Persistent Estrous Rats*

The content of GnRH and its precursor peptide were quantified in female rats bearing lesions in the anterior medial preoptic nucleus (AMPO) and the suprachiasmatic nucleus (SCN), and the effects of the lesions on the synthetic activity of the GnRH neurons were evaluated. Electrolytic lesions which induced persistent estrous (PE), or irregular estrous cycles, were produced by passing 5-10 microA of direct current into the AMPO or the SCN of female rats which exhibited regular 4 days estrous cycles before the lesions. Approximately 5 weeks after lesion placement, blood samples were withdrawn from catheterized, freely moving animals and plasma LH, PRL, estrogen, and progesterone were determined by RIA. The preovulatory surges of LH and PRL were eliminated in AMPO- or SCN-lesioned PE rats. Moreover, the LH surge was eliminated and the PRL surge significantly attenuated after estrogen and progesterone treatment of rats bearing complete lesions, irrespective of the presence of ovaries. Irregular cycling animals with incomplete AMPO or SCN lesions, exhibited attenuated LH surge and PRL surge similar to proestrous controls. In one incidence this occurred spontaneously, and could also be induced by sequential estrogen and progesterone injections. After ovariectomy, plasma LH levels were significantly lower in the lesioned animals as compared to sham operated rats (P less than 0.05). Similar secretory patterns of LH and PRL were obtained from a second series of sham-operated rats during the different stages of the estrous cycle or from AMPO- or SCN-lesioned rats during persistent estrus. After 2 months the animals were killed between 0830 and 0930 h, and the preoptic area and the basal hypothalamus were microdissected from the brain sections. After extraction and purification, proGnRH and GnRH levels were measured by RIA. ProGnRH levels in the preoptic area were significantly reduced in AMPO- or in SCN-lesioned rats, compared to proestrous controls (P less than 0.01). In contrast, GnRH levels in either area did not differ in AMPO- or in SCN-lesioned animals compared to sham-operated, proestrous controls. Therefore, lesions of the AMPO or the SCN produce PE and reduce proGnRH without reducing GnRH levels. These data would suggest that the AMPO and the SCN participate in the control of the estrous cycle and are necessary for preovulatory surges of PRL and LH to occur and that the AMPO and the SCN form part of the neural circuit that regulates GnRH synthesis and/or release.

Induction of persistent diestrus followed by persistent estrus is indicative of delayed maturation of tonic gonadotropin-releasing systems in rats.

The relationship between the amount and duration of administration of estradiol benzoate (EB) to newborn female rats and the induction of sterility was examined in 407 animals. Vaginal smear patterns were classified into 3 types according to the incidence of vaginal proestrus and estrus over a 10-day period: persistent estrous (PE), persistent diestrus (PD), and intermediate (INT), so that the changes in vaginal smear patterns could be analyzed quantitatively. Incidence of the PE pattern was most frequent in the rats that received a single injection of 10 micrograms EB on the day of birth (Day 1). Almost all of the animals receiving 10 daily injections of 10 micrograms EB from Day 1 showed persistent diestrus until at least 100 days of age. In the rats that were given 5 daily injections of 10 micrograms EB Day 1 through Day 5, or a single injection of 100 micrograms EB on Day 3, the incidence of the PD pattern was high at 41-60 days of age, but later the PD-type was replaced by the PE pattern of vaginal smears. In the rats that were treated with 5 daily injections of 10 micrograms EB from Day 1 through Day 5 and were ovariectomized on Day 22, a slight but significant increase in the level of luteinizing hormone in plasma was noted after administration of EB and progesterone on Day 100 but not on Day 50. These results indicated that neonatal injections of EB induce sterility, but the effect is dependent on the amount of EB injected and length of time over which the injections are given.(ABSTRACT TRUNCATED AT 250 WORDS)

Opioid Inhibition of Luteinizing Hormone Release Declines with Age and Acyclicity in Female Rats*

The current study assesses changes in opioid inhibition of LH secretion with age in female rats. We administered naloxone (NAL; 2 mg/kg, iv) to regularly cycling estrous rats of three age groups and measured serum LH in serial samples drawn from intraatrial catheters before and after treatment. The serum LH rise 10 min after NAL treatment in 4- to 6-month-old rats was significantly reduced (P less than 0.01) compared with that in 1.5- to 3-month-old animals, and no LH response was observed in 8- to 11-month-old rats. On early proestrus, LH secretion was also reduced 10 min after NAL treatment in older vs. younger rats, but all groups demonstrated belated LH rises 1 h after treatment during proestrus. Persistent estrous (PE) rats released less LH after NAL treatment than age-matched estrous rats (P less than 0.025). Higher dose NAL treatment did not increase LH release in estrous or PE rats. These results support the hypothesis that opioid inhibition of LH secretion diminishes with age in cycling rats. Furthermore, opioid tone is a function of estrous state as well as age. PE rats have lower opioid tone than cycling animals of the same age. Our findings suggest a possible role of diminished opiate tone in reproductive senescence.

Transmission and scanning electron microscopic examination of rat vaginal epithelium in the normal estrous cycle and in persistent estrous

AbstractThe rat vaginal epithelium in both normal and persistent estrous (PE) cycles was examined by transmission and scanning electron microscopy utilizing cardiac perfusion and ruthenium red staining. It was demonstrated that surface vaginal epithelium in persistent estrous exhibited irregularly shaped cells. Borders were not well defined as is the case in normal estrous, and cells lacked the complex, interconnecting microridges and pits. Vaginal surfaces were completely smooth or covered with few microvilli. In the control specimens the fine structure en block stained with ruthenium red showed distinct cell membranes bordering the intercellular spaces between epithelial cells. With PE rats, this finding was absent, and there was instead a significant expansion of intercellular space with villiform processes of the cytoplasm projecting into the space. Utilization of the combination of techniques described provided the investigators with a better definition of vaginal tissue ultrastructure and revealed significant morphological differences between the normal estrous cycle and persistent estrous in vaginal specimens from rat models.

Cervical and vaginal surface epithelial changes in persistent estrous, chemically diabetic BB wistar rats observed by scanning electron microscope

Studies of the effects of diabetes on reproduction in the female BB Wistar rat are not available in the literature. Male reproduction has been studied in these spontaneously-diabetic BB Wistar rats, and abnormalities of the reproductive tract have been found. This investigation, therefore, compared the ultrastructure of the lower reproductive tract of the chemically-diabetic female BB Wistar rat which has developed persistent estrous(PE) to normalcycling controls.

Surface Changes in Vaginal Epithelial Cells During Persistent Estrous Rats

Light exerts a major influence on the reproductive functions of the female rat, and the presence of constant light results in an aberrant vaginal cycle, leading to infertility associated with polycystic ovary (PCO) syndrome. In the present communication, we have utilized scanning electron microscopy (SEM) to study changes on the surface epithelium of persistent estrous (PE) rats with PCO.Sprague-Dawely female rats were exposed to constant light for 50-150 days according to protocol that has been previously described. Rats in normal estrous, as determined by vaginal smears, were used as controls. Nembutal- anesthesized rats were perfused through the aorta with 2.5% gluteraldehyde in 0.1M sodium cacodylate buffer (pH 7.3). The mucosa of the upper third of the vagina from PE and control rats were microdissected, processed for SEM studies by a modified 0T0T0 technique, dehydrated, and critical point dried with C02- The specimens were examined in a Hitachi scanning electron microscope Model S-500.

Relationship between circulating estrogens and the central mechanisms by which ovarian steroids stimulate luteinizing hormone secretion in aged and young female rats.

The present study was undertaken to determine the relationship between a persistent elevation in circulating estrogens and the lack of a stimulatory feedback of ovarian steroids on LH secretion in old persistent estrous (PE) rats and to investigate whether, under experimental conditions in young cyclic females, a chronic rise in circulating estradiol inhibits the positive feedback action of steroids on LH secretion. Eighteenmonth-old PE rats and 4-month-old regularly cyclic females were ovariectomized (OVX) and immediately primed with estradiol benzoate (EB; 8 /μg/100 g BW, sc). Three days later, progesterone injection (0.5 mg/100 g BW, sc) elicited a LH surge in formerly cyclic but not PE rats. Five weeks later, however, EBprogesterone administration to the same animals elicited a LH surge not only in OVX young rats but also in OVX old females which had previously been in the PE state, although the magnitude of the LH surge was significantly less in OVX-PE than in OVX young rats. In long term OVX old and...

Ultrastructural observations of granule extrusion from rat anterior pituitary cells after injection of LH-releasing hormone.

In order to observe the ultrastructural aspects of granule extrusion from gonadotrophs, the authors injected pure, natural porcine LH-releasing hormone (LH-RH) into persistent-estrous (PE) rats. Clear-cut extrusions of secretory granules appeared from LH-gonadotrophs 2.5, 5 and 15 minutes after the injection. Massive extrusion was observed at 15 minutes. Serum LH and FSH were also estimated by radioimmunoassay on blood samples taken at the same times after the injections. LH was increased in the blood sera after the injection of LH-RH, but serum FSH was not significantly different among the various treated rats. The rise in serum LH after LH-RH injection was well correlated with the ultrastructural phenomenon of granule extrusion from LH-gonadotrophs.

Spontaneous activity of neonatally estrogenized female rats.

The running activity of normal and neonatally estrogenized persistentestrous (PE) and-diestrous (PD) rats was investigated by recording the number of revolutions of the drum attached to the activity cage, under a controlled photoperiod (14 hrs. L: 10 hrs. D). Normal rats displayed 4-day cyclic changes in spontaneous activity, while PE rats was acyclic. PD rats were also acyclic and definitely lessactive than normal and PE rats. Ovariectomy resulted in a drop in activity in normal and PE rats, although the decrease was less marked in PE rats than in normal rats. Ovariectomy brought about any perceptible changes in activity of PD rats. Following injections of 10μg estradiol-17β for 7 days, spontaneous activity was increased in normal and PE rats but not in PD rats. The increase was greaterin magnitude in normal than in PE rats. These findings indicate that PE rats is less sensitive to rise and fall in estrogen titer in the circulating blood. Daily injections of 100μg estradiol-17β evoked an increase in activity of normal and PE rats, approximately similar in magnitude to that following injections of 10μg estradiol. Activity of PD rats was also increased after injections of 100μg estradiol. However, difference in response to estradiol between normal and PD rats was highly significant. It is concluded that neural areas involved in regulation of spontaneous activity is permanently suppressed by neonatal estrogenization.

Pituitary-ovarian function in rats exposed to constant light: a chronological study.

A chronological study was made of adenohypophyseal, ovarian, uterine, and vaginal function in 2 groups of adult rats exposed to continuous light (LL): (I) those which were still showing vaginal smear cycles, and which were autopsied during proestrus and estrus of cycles 1–5 in LL, and (II) those which had shown 5, 10, 15, 20 or 30 days of persistent vaginal estrus. Ovulation was still occurring in Group I; furthermore, the ovulatory surge of LH was being released at a time of day similar to that in animals exposed to a light-dark (LD) regimen. However, the exposure to LL did appear to lead to some desynchronization of 24-hr rhythms within the hypothalamic-pituitaryovarian axis; a persistent-estrous (PE) state (Group II) eventually developed, characterized by a failure to release an ovulatory surge of LH and by a “steady-state” secretion of ovarian steroids and folliculotrophin (i.e., “tonic” FSH and LH). The PE state, in turn, could be subdivided into 2 separate stages as a function of the number of days ...