Delay In Vaginal Opening Research Articles

EtOH disrupts female mammalian puberty: age and opiate dependence.

The major drug of abuse among teenagers in the United States continues to be ethanol (EtOH), but use is seen in children as young as nine. In the studies reported here, the impact of EtOH on biologic and hormonal parameters of puberty was assessed in female rats. Rats were fed a liquid diet containing EtOH, pair fed an identical liquid diet containing dextrimaltose instead of EtOH, or fed a liquid diet not containing EtOH ad libitum. Feeding was started at 21, 25, or 28 d of age. EtOH markedly delayed the age at vaginal opening (34.5 +/- 0.5 d in controls vs 48.5 +/- 2.4 d in EtOH animals; p < 0.001), delayed the age at first estrous (40.9 +/- 0.6 d in controls vs 61.2 +/- 2.6 d in EtOH animals; p < 0.001), increased the length of the estrous cycle, and decreased the number of proestrous days. EtOH, concomitant with reduced ovarian and uterine weight, decreased serum estradiol and progesterone. Associated with these changes in ovarian hormones there was a selective increase in follicle-stimulating hormone, but not luteinizing hormone. EtOH consistently reduced insulin-like growth factor-1. In general, EtOH-induced disruption was more severe the younger the animals were at the start of feeding. Opiate receptor blockade with naltrexone completely prevented the EtOH-induced delay in vaginal opening. The impact of EtOH on female puberty is dramatic, is an emerging public health problem, and deserves more study.

TWO-GENERATION REPRODUCTIVE TOXICITY STUDY OF TRIBUTYLTIN CHLORIDE IN FEMALE RATS

A two-generation reproductive toxicity study of the effects of tributyltin chloride (TBTCl) was conducted in female rats using dietary concentrations of 5, 25, and 125 ppm TBTCl. Reproductive outcomes of dams (number and body weight of pups and the percentage of live pups) and the growth of female pups (the day of eye opening and body weight gain) were significantly decreased in the 125 ppm TBTCl group. A delay in vaginal opening and impaired estrous cyclicity were also observed in the 125 ppm TBTCl group. However, an increase in anogenital distance was found in all TBTCl groups on postnatal d 1. A dose-effect relationship was observed in TBTCl-induced changes in anogenital distance. These results indicate that the whole-life exposure to TBTCl affects the sexual development and reproductive function of female rats. In addition, the TBTCl-induced increase in anogenital distance seems to suggest it may exert a masculinizing effect on female neonates. However, the concentrations of TBTCl used in this study are not environmentally relevant.

The effects of atrazine on female wistar rats: an evaluation of the protocol for assessing pubertal development and thyroid function.

The effects of atrazine (ATR), a chlorotriazine herbicide, on the onset of puberty were evaluated in Wistar rats. Female rats were dosed by oral gavage from postnatal day(s) (PND) 22 through PND 41 with 0, 12.5, 25, 50, 100, or 200 mg ATR/kg. Vaginal opening (VO) was significantly delayed 3.4, 4.5, or greater than 6.8 days by 50, 100, and 200 mg/kg, respectively. VO had not occurred in 4 of 15 females in the 200 mg/kg group by the time of necropsies (PND 41). Body weight (bw) at necropsy was reduced in the 200 mg/kg group by 11.6%, but was not different from the control (0) in the 50 and 100 mg/kg groups. To examine the influence of reduced bw on pubertal development, a group of pair-fed controls was included whose daily food intake was dependent upon the amount consumed by their counterpart in the 200 mg/kg group. Although necropsy bw was reduced to the same extent as the ATR females, VO in the pair-fed controls was not significantly delayed. Adrenal, kidney, pituitary, ovary, and uterine weights were reduced by 200 mg/kg ATR. Serum T(3), T(4), and TSH were unaltered by ATR, which was consistent with no histopathologic/morphologic changes in the thyroid. Estrous cyclicity was monitored in a second group of females from VO to PND 149. The number of females displaying regular 4- or 5-day estrous cycles during the first 15-day interval after VO was lower in the 100 and 200 mg/kg ATR and pair-fed controls. Irregular cycles were characterized by extended periods of diestrus. By the end of the second 15-day interval (PND 57-71), no effects on estrous cyclicity were observed. These data show that ATR can delay the onset of puberty and alter estrous cyclicity in the female Wistar rat ( NOAEL of 25 mg/kg). Reduced food consumption and bw did not account for the delay in VO, because this effect was not observed in the pair-fed controls. In addition, the effect on estrous cyclicity was observed in the 100 mg/kg ATR group where no significant reduction in bw was observed.

Alpha-difluoromethylornithine modifies FSH secretion and puberty onset in the female rat.

FSH secretion is high in immature female rats from Postnatal Day 5 to 18 and decreases thereafter. This is a relatively steroid-independent event of cerebral origin and of importance for puberty onset. Polyamines, a group of ubiquitous amines, play an essential role in tissue growth and differentiation, body weight increment, brain organization, and molecular mechanisms of hormonal action. Polyamine levels as well as the activity of ornithine decarboxylase, the limiting enzyme in polyamines biosynthesis, are highest during development. Inhibition of their synthesis during this period by alpha- difluoromethylornithine (DFMO), a specific and irreversible inhibitor of ornithine decarboxylase, impairs normal brain development. The present study tested the hypothesis that polyamines play a role during brain organization of reproduction. DFMO was administered following different schedules in female newborn rats, and the effect on pituitary secretion, puberty onset, and fertility was evaluated. In three groups (daily injections from Day 1 to 9, or from Day 1 to 6, or injections on alternative days from Day 1 to 9), a delay in vaginal opening and first estrous was observed. When vaginal opening was plotted against body weight, it was evident that in groups daily injected with DFMO vaginal opening occurred at a lower body weight. In the group treated on alternate days, a delay occurred but at a higher body weight than in controls. In this group, serum FSH levels on Day 10 and 20, but not on Day 30, were higher in DFMO rats. In the group treated from Day 1 to 6 daily, DFMO increased serum FSH on Postnatal Day 20. After vaginal opening, estrous cyclicity in control and DFMO injected rats was similar. There was no significant effect of treatment on fertility and litter weight or number of offspring at birth. It is concluded that DFMO, an inhibitor of ornithine decarboxylase, administered during the first week of life in female rats is followed by prolonged high FSH serum levels and delayed puberty, but once puberty occurs, fertility is normal.

Temporal Energy Restriction and the Timing of Vaginal Opening in Female Rats.

The effect of a temporary dietary restriction (10 g/day) duirng gestation or lactation on the subsequent maturation of female offspring was examined. Mother rats were divided into 6 groups according to the period of restriction: namely, Group A (between days 0 and 7 of gestation), Group B (between days 8-14 of gestation), Group C (between days 15-21 of gestation), Group D (between days 1-7 of lactation), Groupt E (between days 8-14 of lactation), Group F (no restriction). In Group E, the mother's body weight decreased by 17% compared to that on day 0 of gestation. Female pups in this group showed retardation in growth rate concurrent with a delay in vaginal opening. No significant effects were found in any other groups. These results suggest that adequate nutrition during midlactation is critical for optimal growth of female pups and subsequent opening of the vagina.

Delay of puberty and impairment of growth in female rats given a non competitive antagonist of NMDA receptors

Reportedly, excitatory amino acids are involved in the control of gonadotropin secretion of rats and non-human primates. The aim of this study was to investigate the effect of chronic blockade of NMDA (N-methyl-D-aspartic acid) receptors by the non competitive receptor antagonist MK-801 on gonadotropin secretion and the onset of puberty in female rats. Moreover, since in humans alterations of the timing of puberty frequently coexist with disturbances of body growth, suggesting a common etiology for both events, we evaluated the effect of MK-801 also on the neural mechanisms controlling growth hormone (GH) secretion. Twenty-one-day-old female rats were treated with MK-801 (0.2 mg/kg ip, bid) or placebo for 10 days and were killed after 7 days of withdrawal. Administration of MK-801 induced a significant impairment of growth rate without altering food intake, and a delay in vaginal opening. Pituitaries from rats treated with MK-801 had a reduced luteinizing hormone (LH) content, and secreted in vitro lower amounts of LH both under basal and LHRH-stimulated conditions. MK-801 treated rats had a lower pituitary GH content and basal and GHRH-stimulated GH release and reduced plasma insulin-like growth factor-I levels. These data indicate that blockade of NMDA receptors in a critical period of the female rat life-span: 1) delays puberty by reducing gonadotropin secretion; 2) impairs growth rate by reducing GH secretion, with a mechanism still to be clarified.

Seizure-induced delay of puberty in female rats: effects of age, stress and opioid antagonists.

We have shown that pre- and post-pubertal female rats are sensitive to seizures. For example, daily convulsions commencing at 24 days of age delay puberty. Here we examine the effect of seizures at various ages. In addition, because opioid peptides are implicated in regulating the onset of puberty and are activated by convulsions, we also investigate the effect of opioid antagonists in the seizure-induced delay of puberty. A single daily electroconvulsive shock (ECS) was given for 10 days to neonatal (days 2-11), infantile (days 15-24) and juvenile (days 22-31) rats. The treatment delayed vaginal opening (VO) in juvenile rats. Neonatal and infantile rats were unaffected. VO was also delayed by daily ECS for only 5 days in the late juvenile (days 27-31) period. The opioid receptor antagonists naloxone, naltrexone and nalmefene injected before and after single daily ECS were unable to block this effect of ECS on VO. To examine whether the effect of ECS is related to stress, we examined several stressors known to induce opioid-mediated alterations in gonadotrophin secretion. Footshock, immobilization and ether stress administered in the juvenile period (days 27-31) did not affect the timing of VO. In addition, rats anaesthetized with halothane, and then given ECS, still showed a delay of VO. These data demonstrate that rats in the late juvenile stage of development are most sensitive to convulsions. We also suggest that opioids are not critical to the mechanism by which the ECS disturbs puberty, and that ECS elicits its effect seemingly independently of the convulsive stress.

Possible antiestrogenic activity of lindane in female rats

During chronic peroral (PO) treatment of weanling, female Fischer 344 rats with daily injections (0.069 mmol/kg) of either 1,1'-(2,2,2-trichloroethylidene) bis [4-chlorobenzene] (p,p'-DDT), 2,4-dichlorophenoxy acetic acid (2,4-D), or gamma-hexachlorocyclohexane (lindane), the lindane treatment induced a significant 20% increase in body weight after 110 days. Further investigation with 0, 5, 10, 20, and 40 mg/kg lindane confirmed a significant increase in average body weight gain at the two highest doses after ten weeks of treatment. Significantly greater food consumption was observed, and the Lee index indicated that lindane treatment induced obesity. In addition to obesity, lindane caused a delay in vaginal opening, disrupted estrous cycling, reduced pituitary and uterine weight, and elevated food consumption during proestrus (when appetite is normally suppressed by estradiol). These responses suggest that, by inducing alterations in the reproductive function of the female rat and by interfering with hormonal regulation of energy balance, lindane may be antiestrogenic rather than estrogenic as previously proposed.

The effects of perinatal exposure to nicotine on plasma LH levels in prepubertal rats

Adult female rats were chronically treated with nicotine administered via the drinking water during pregnancy and/or lactation. The approximate doses of nicotine consumed per day were 2.4 mg/kg and 4.5 mg/kg of body weight. The pups were weaned at 20 days of age. The pups were killed by decapitation on postnatal days 20, 30, or 40 and plasma from heparinized trunk blood was assayed for luteinizing hormone (LH). At 30 days of age untreated male and female offspring had the highest levels of plasma LH compared to 20 and 40 days of age. This level was not affected by any subsequent dose or treatment. Prepubertal females exposed to nicotine during pregnancy failed to exhibit the pattern of LH levels seen in control animals, whereas those exposed during lactation or throughout the perinatal period showed a distinctive pattern of plasma LH. Chronic exposure of female offspring to the low dose of nicotine during lactation tended to increase plasma LH levels at 20 and 40 days. Female offspring exposed to nicotine during pregnancy or to the low dose during lactation showed significant deficits in body weight at 40 days of age which appeared to correlate with a delay in vaginal opening. The results suggest that perinatal exposure to maternally administered nicotine may disrupt normal patterns of LH release in the offspring of both sexes and alter sexual development in female offspring.

The Combined Effects of Dietary Protein and Fat on Prolactin in Female Rats

A 3 × 3 factorial experiment was conducted to examine how protein content (8, 16 and 32% of kcals from casein) and fat content (12, 24 and 48% of kcals from corn oil) interact to influence prolactin homeostasis in female rats. Weanling Sprague-Dawley rats were assigned to each of nine diets fed ad libitum. Dietary fat had no effect on the time of vaginal opening although first estrus occurred slightly earlier with high fat diets (P < 0.05). Rats fed diets with protein at 8% of kcal showed a 2-wk delay in vaginal opening and first estrus. After 18–20 wk of feeding, serum and pituitary prolactin concentrations were determined after decapitation during the afternoon of proestrus or diestrus. Although serum prolactin was elevated at proestrus, there were no effects of fat or protein on the serum or pituitary prolactin. Another group of rats were fitted with jugular cannulae and treated with perphenazine, a potent prolactin-stimulating drug. Dietary fat or protein had no effect on prolactin secretion after perphenazine treatment. In the final experiment, rats fed each diet were treated with either i.p. saline or perphenazine followed by decapitation. The pituitary was removed 3 h later for determination of the amount of prolactin depleted from the pituitary. There was no effect of diet on the amount of pituitary prolactin depleted by perphenazine. No evidence was obtained that changes in dietary fat or protein concentration influence serum prolactin or its secretion in response to a provocative stimulus. This study supports the hypothesis that dietary fat enhances mammary carcinogenesis by mechanisms independent of prolactin concentrations in the serum and pituitary prolactin secretion. Previous studies showing an effect of fat on serum prolactin may have been due to a deficiency of essential fatty acids.

Paradoxical Effects of D-TRP6-LHRH in Immature Female Rats Correlated with Changes in ACTH, Prolactin, and Corticosterone Levels1

Immature female rats were treated with D-Trp6-LHRH to determine if the delay in vaginal opening induced by the superactive agonist of LHRH could be correlated with changes in ACTH, prolactin, or corticosterone levels. The rats were injected daily from Day 30 to Day 40 of age with 0.05 or 1 μg of D-Trp6-LHRH or with saline alone. Six animals per group were sacrificed on Days 31, 35, 38, or 42 and trunk blood collected. Pituitary and adrenal weights did not vary between groups, but increased in all treatment regimens as the animals grew older. Prolactin and ACTH levels increased on Days 35 and 38 in the control animals, but were significantly reduced in both agonist-treated groups. Relative to controls, serum corticosterone was elevated (P<0.05) on Day 31 in 0.05 μg-treated animals and decreased on Day 38 in animals treated with 1 μg D-Trp6-LHRH. The addition of corticosterone (160 μg/ml in 4% ethanol) to the drinking water reversed the inhibitory effects of 1 μg of D-Trp6-LHRH on vaginal opening. In animals treated with saline or 0.05 μg D-Trp6-LHRH, corticosterone did not affect the normal vaginal opening. These results demonstrate that it is possible that D-Trp6-LHRH delays vaginal opening through its effect on corticosterone levels.

Intrinsic neurons of the septal area are involved in reproductive development of the female rat. Alterations after kainic acid lesioning.

Three experiments were conducted using intracranial infusion of kainic acid (KA) to lesion the medial septal (MS) neurons and dorsal hippocampal (DH) neurons in 25-day-old female rats. Rats of experiment 1 were allowed to develop until vaginal opening (VO) after which vaginal lavages were taken to monitor the vaginal cycle. Rats of experiment 2 received estradiol benzoate (EB) subcutaneously immediately after KA infusion. Blood was obtained at 11.00 and 16.00 h, 2 days after EB and measured for luteinizing hormone (LH) by radioimmunoassay. Rats of experiment 3 received MS KA lesions and 2 days later tested for pituitary responsiveness to LHRH-induced LH release. In experiment 1, MS KA-lesioned rats exhibited a significant delay in VO when compared to the sham-lesioned or the unoperated control groups. Hippocampal KA-lesioned rats in contrast, showed no difference in the age of VO from the control groups. Septal KA-lesioned rats also showed disrupted vaginal cycles after the onset of puberty. In experiment 2, all the EB-treated animals exhibited the expected rise in serum LH on the afternoon 2 days following EB with the exception of the septal KA-lesioned rats. In every case, KA infusion into the medial septum abolished the ability of EB to induce an LH increase at 16.00 h 2 days after EB administration. DH KA lesion on the other hand had no effect on the EB-induced LH increase. The block of LH secretion by MS KA infusion shown in experiment 2 is not a result of decreased pituitary gland sensitivity to LH-releasing hormone (LHRH) as shown in experiment 3. These results indicate that MS neurons play a vital role in the feedback mechanism whereby EB induces an LH increase in the prepuberal female rat, and that they play a role in the timing of VO.

Monosodium glutamate disruption of behavioral and endocrine function in the female rat.

Experiments were conducted to determine the effects of neonatal administration of L-monosodium glutamate (MSG) on behavioral and endocrine function in the female rat. Administration of MSG (4 mg/kg body weight) at days 1, 3, 5, 7 and 9 in neonates results in a delay of vaginal opening (VO) and the absence of ovulation at the time of VO. However, some rats were observed to ovulate after VO if they were subjected to sequential laparotomies. MSG-treated rats also fail to exhibit compensatory ovarian hypertrophy. Ovariectomized MSG-treated rats injected with estradiol benzoate (EB) followed by a progesterone injected 2 days later did not exhibit sexual beahvior to male rats, while all the control rats displayed lordosis. Chronic treatment with EB for 12 days, followed by a progesterone injection on the 12th day, resulted in a marked improvement of the sexual receptivity of the MSG-treated rats. The body weight of the MSG-treated animals was lower than that of the controls during development although the MSG animals looked obese. Food intake is normal in the MSG-treated rats, but when expressed as intake/100 g body weight, the MSG-treated rats appeared slightly hyperphagic, MSG-treated rats respond with increased food intake after ovariectomy and EB treatment suppresses the increased food intake. Thus, the control of food intake by estrogen does not seem to be affected by the MSG treatment; in fact, these animals seem to be more sensitive than control rats to the anorectic effects of EB. Neonatal MSG treatment appears to affect the neural control for the tonic secretion of gonadotropins by destroying arcuate nuclei. This undoubtedly reduces the reproductive capacity of the animals by impeding the growth and secretions of their ovaries. The findings that chronic estrogen followed by progesterone treatment can reinstate sexual receptivity in MSG-treated animals suggests that the arcuate nuclei are not needed for the expression of sexual behavior and that estrogens might remedy the fertility problems of MSG-treated animals.

Paradoxical Effects of d-Trp6-Luteinizing Hormone-Releasing Hormone on the Hypothalamic-Pituitary-Gonadal Axis in Immature Female Rats

The effect of administration of a superactive and long-acting analog of luteinizing hormone-releasing hormone (LH-RH), D-Trp6-LH-RH, in doses of 0.05 or 1 microgram/day for 10 days on the hypothalamic-pituitary-gonadal axis was studied in immature female rats. Treatment with a 0.05-microgram dose of analog produced few changes as compared with the control group. Treatment with 1 microgram of D-Trp6-LH-RH did not affect the body weight or the pituitary weight, but increased ovarian weight and decreased uterine weight; elevated serum gonadotropin levels; and lowered the pituitary LH content. This depletion of pituitary LH content was associated with a low pituitary responsiveness to LH-RH. Serum estradiol levels were not modified, suggesting that decreased uterine weight reflects a direct and extrapituitary effect of this analog. The hypothalamic LH-RH content was higher, indicating a possible inhibition of the release of endogenous LH-RH. A delay in vaginal opening was also observed. This indicates that large doses of D-Trp6-LH-RH may interfere with the process of puberty in rats. These findings extend other reports about the paradoxical antifertility effects of large doses of stimulatory analogs of LH-RH.

Neonatal ablation of the suprachiasmatic nucleus. Effects on the development of the pituitary-gonadal axis in the female rat.

Ablation of the suprachiasmatic nucleus (SCN) at 2 days of age, prior to the formation of the retinohypothalamic projection, produces a permanent state of constant vaginal estrus in the postpubertal female rat. Although such lesions do not alter the onset of puberty in sighted rats, they do compensate for the delay in vaginal opening induced by neonatal binding. The ovaries of sighted and blinded SCN-lesion rats are small and polyfollicular and the pituitaries of blinded SCN-lesion rats are abnormally large. Sampling of plasma in the morning and afternoon for up to 12 consecutive days in sighted SCN-lesion rats reveals continuously low luteinizing hormone levels. This constellation of endocrine alterations does not correlate with damage to any structures outside the SCN. Since the organization of the rodent estrous cycle is circadian, these results emphasize further the importance of the SCN in circadian rhythm generation. The necessity of an intact SCN for the development of normal, cyclic reproductive function implies that sparing or recovery of function does not occur.

Adrenal involvement in the timing of puberty in female rats: interaction with serum prolactin levels.

ABSTRACT Female Sprague-Dawley derived rats were either sham-operated or adrenalectomized at 20 days of age. Treatment of separate groups of rats with progesterone, corticosterone or prolactin was started at 21 days of age, and was continued until the day of vaginal opening. Similarly treated rats were decapitated at 25, 29 and 33 days of age and at the day of vaginal opening and the sera were collected from each animal and assayed for prolactin, LH and FSH. All rats were decapitated between 4–6 p. m. Adrenalectomy, as previously reported, significantly delayed the day of vaginal opening. Prolactin treatment advanced the day of vaginal opening in intact rats and it restored the day of vaginal opening in adrenalectomized rats to that of intact control rats. Progesterone treatment always delayed the day of vaginal opening in intact rats. It caused an even greater delay in vaginal opening than adrenalectomy alone. Corticosterone treatment, on the other hand, was able to reverse the effect of adrenalectomy but had no effect on vaginal opening in intact rats. In intact rats serum prolactin levels rose steadily until the day of vaginal opening. Adrenalectomized and intact rats treated with progesterone had significantly decreased serum prolactin levels. However, in adrenalectomized corticosterone treated rats serum prolactin levels were significantly increased at all ages measured when compared to the untreated rats. The FSH and LH levels showed no significant changes during any of these experimental procedures. Approximately 50 % of the rats showed LH peaks on the day of vaginal opening as well as an increase in FSH. The data indicate a prolactin-adrenal interaction for the timing of the onset of puberty in immature rats, assuming that the day of vaginal opening in association with increased gonadotrophin levels is an indicator for the occurrence of puberty.

Serum Prolactin Concentrations in the Adrenalectomized Rat: Relationships to Puberty Onset

The purpose of this study was to examine the hypothesis that prolactin is involved in puberty onset by comparing changes in serum prolactin (PRL) concentrations between weaning age and puberty in normal rats and in rats in which puberty was delayed by means of adrenalectomy at 22 days of age. In intact rats a consistent elevation in serum PRL was observed between 28 and 30 days of age (a rise from a mean of 8.4 ±1.4 ng/ml PRL to 24.6 ± 2.4 ng/ml PRL). After adrenalectomy at 22 days of age there was a depression of PRL lasting several hours. This was followed by an elevation which persisted for the next 48 h. The PRL elevation observed in intact rats between days 28–30 was absent in rats adrenalectomized at 22 days of age. Associated with the absence of the elevation in serum PRL, there was a delay in vaginal opening and ovulation of 4 to 5 days. Replacement therapy with corticosterone in the drinking water (80 μg/ml) resulted in normal vaginal opening and ovulation times but failed to restore the prolacti...

The Inhibitory Effects of ACTH and Adrenalectomy on Reproductive Maturation in Female Rats

The effects of ACTH and adrenalectomy on sexual maturation was studied in female rats. Weanling (21-day-old) intact and adrenalectomized female rats received 4 IU/100 gm body weight until Day 40. Animals were sacrificed on Days 25 30 35 40 and on the day of vaginal opening. Blood samples were assayed for progesterone corticosterone luteinizing hormone (LH) and prolactin. Animals sacrificed at 1st estrus were examined for the number of ova present in the Fallopian tubes. ACTH treatment delayed vaginal opening for 4 days in both intact and adrenalectomized animals. Since ACTH-injected adrenalectomized rats did not show a delay in vaginal opening it would appear that the inhibitory effect of ACTH is mediated by the adrenal glands. However these animals had a marked reduction in the number of ova present at 1st estrus and markedly lower body weight at vaginal opening. ACTH-treated intact rats that showed delayed vaginal opening had elevated levels of progesterone and corticosterone. In animals not exhibiting retarded sexual maturation only corticosterone levels were elevated. Although it appeared that the chronically elevated levels of progesterone in ACTH-treated rats inhibited sexual maturation by interfering with plasma LH attempts to show a suppression of plasma LH were unsuccessful.

Delay of Puberty and Inhibition of Reproductive Processes in the rat by a Gonadotropin-Releasing Hormone Agonist Analog

Studies were performed in immature and mature female rats to determine the effects of chronic administration of [D-Leu6,des-Gly-NH210, Pro-ethylamide9]-gonadotropin-releasing hormone (GnRH) upon maturation and maintenance of reproductive function. Treatment of immature rats inhibited normal overian growth and maturation as evidenced by arrest of uterine growth, delay of vaginal opening, and absence of normal cycling. The primary responses in the mature animal were cessation of cycling, atrophy of the ovaries to a "prepuberal" size, and uterine weight regression. In both groups termination of treatment was followed by prompt restoration of normal ovarian mass and function, including support of breeding processes. The results presented here suggest that chronic administration of the GnRH analog inhibits ovarian steroid output either by direct action of the analog on the ovary or indirectly through qualitative or quantitative changes in levels of luteinizing hormone and follicle-stimulating hormone.

The effect of pineal methoxyindoles on rat vaginal opening time.

SUMMARY Melatonin and 5-methoxytryptophol, the two methoxyindoles of pineal origin, were injected into a lateral ventricle of the brain of immature female rats. Treatment was started on the 25th day of age and terminated when the vagina opened. The injection of both methoxyindoles resulted in a statistically significant delay in vaginal opening. Since previous experiments had shown that melatonin specifically inhibits secretion of luteinizing hormone and that 5-methoxytryptophol specifically blocks release of follicle-stimulating hormone, the present results support the hypothesis that the onset of sexual maturation needs a balanced secretion of both gonadotrophins.