Postnatal profile of plasma leptin concentrations in male and female rats: relation with the maturation of the pituitary-gonadal axis

Leptin, the ob gene product, has been implicated in the initiation of puberty in mice and humans. However, it is not yet clear whether leptin also plays a pivotal role in promoting sexual maturation in rats. Based on the assumption that circulating leptin levels would increase during the peripubertal period if this hormone triggers puberty, we examined the developmental profile of plasma leptin from neonatal (day 1) through adult (day 85) age in both male and female rats and simultaneously monitored several important indices of pituitary-gonadal function. A significant elevation of plasma leptin during the peripubertal period was not observed in either male or female rats. Although this finding may not support leptin as a humoral factor triggering puberty, we observed a rise in leptin in both sexes from the second postnatal week, which clearly preceded the first significant elevations of luteinizing hormone and gonadal steroids. Therefore, it is still possible that leptin may play a role in promoting sexual maturation in rats of both sexes already from an early postnatal period. It seems that the role for leptin in sexual development is permissive, but not decisive, in rats.     

Role of early experience in social behaviour of laboratory-bred female rats

Social relationships among female laboratory-bred rats in a community are influenced by their early life history. When the rats were born and kept until adulthood under conventional breeding conditions, i.e. in single cages, and then used to form a community in which they gave birth, one female assumed the dominant role, with all others subordinate. The dominant female herded all young rats born in the community, irrespective of their age, in a single litter and suckled them. She accumulated food and wood shavings from other parts of the community near the nest and prevented access to the nest to all other community members including other females that had given birth. Subordinate females ceased to show maternal behaviour, including lactation, within 24 h (occasionally within 48 h). The mortality of the young until 15 days of age was high. This type of behaviour in a community was observed both with randomly chosen female rats and with rats selected as dominant and subordinate types in preceding experiments. Female rats born and reared in a community and rats living in a community from 15 to 30 days of age did not differentiate into dominant and subordinate types. All females retained their maternal behaviour, including lactation. Mortality of young rats was minimal. In most cases the females built one common nest; sometimes each female built her own nest. The results point to the decisive role of early experience in the development of maternal behaviour and in the occurrence of communal rearing of the young.     

Effects of gonadotropin-releasing hormone administration on the pituitary-gonadal axis in male and female dogs before and after gonadectomy

GnRH-stimulation tests were performed in 14 female and 14 male client-owned dogs of several breeds, before and 4 to 5 mo after gonadectomy. The aim of the study was to obtain more insight into the pituitary-gonadal axis in intact and neutered dogs and to establish reference values. Basal plasma luteinizing hormone (LH) and follicle-stimulating hormone (FSH) concentrations were increased significantly after gonadectomy in both bitches and male dogs. In both males and females ranges of the basal plasma FSH concentrations, before and after gonadectomy, did not overlap as opposed to the overlap in ranges of the basal plasma LH concentrations. Before gonadectomy basal plasma LH concentrations were lower and basal plasma FSH concentrations were higher in bitches than in male dogs. After gonadectomy these basal values did not differ significantly. GnRH administration before gonadectomy resulted in an increase in plasma LH and FSH concentrations in both genders. GnRH administration after gonadectomy produced an increase only in plasma LH concentrations in both genders, and a just significant increase in plasma FSH in castrated male dogs. GnRH administration before gonadectomy resulted in a significant increase in plasma testosterone concentration in both genders. In males ranges of basal and GnRH-stimulated plasma testosterone concentrations before and after gonadectomy did not overlap. Basal plasma estradiol concentrations were significantly higher in intact males than in castrated males and their ranges did not overlap. The basal estradiol concentrations in bitches before and after ovariectomy were not significantly different. At 120 min after GnRH administration, ranges of plasma estradiol concentration of intact and ovariectomized bitches no longer overlapped. In conclusion, basal plasma FSH concentration appears to be more reliable than basal plasma LH concentration for verification of neuter status in both male and female dogs. The basal plasma testosterone concentration appears to be reliable for verification of neuter status in male dogs. The plasma estradiol concentration at 120 min after GnRH administration can be used to discriminate between bitches with and without functional ovarian tissue.     

芹菜素对雌性大鼠生殖轴的影响

目的：探讨芹菜素在10-9mol/L浓度时对雌性大鼠生殖功能的影响。方法：应用侧脑室注射方法观察芹菜素对雌性大鼠生殖轴激素含量的影响。在侧脑室注射后第3天取血浆,采用放免技术测定血浆中促性腺激素释放激素（GnRH）、卵泡刺激素（FSH）、黄体生成素（LH）、雌二醇（E2）、孕酮（P）的含量。结果：在给药后第3天血浆中促性腺激素释放激素（GnRH）、卵泡刺激素（FSH）、黄体生成素（LH）的含量增加而雌二醇（E2）、孕酮（P）的含量降低,差异有显著性。结论：芹菜素抑制雌二醇（E2）合成中芳香化酶的活性实现对雌二醇（E2）、孕酮（P）分泌的抑制。芹菜素通过影响雌激素受体而使下丘脑的促性腺激素释放激素（GnRH）和腺垂体的卵泡刺激素（FSH）、黄体生成素（LH）分泌增加。     

Effect of pentoxifylline on endothelaemia and hypothalamic-pituitary-adrenocortical axis activation in female rats under stress exposure

Endothelial dysfunction may belong to negative consequences of stress exposure accompanied by activation of several stress systems including the hypothalamic-pituitary-adrenocortical (HPA) axis. The present experiments were aimed at testing the hypotheses that i) immobilization (IMO) stress results in sustained increase in endothelaemia for 24 h and that ii) pentoxifylline, a drug with endothelium protective properties, attenuates the rise in endothelaemia and HPA axis activation in female rats as shown previously in males. Circulating endothelial cells increased immediately after the IMO for 2 h, returned back to control levels at 12 h and increased again at 24 h. Stress-induced rise in adrenocorticotropic hormone (ACTH) and corticosterone levels was particularly high immediately after the IMO. Pretreatment with pentoxifylline (20 mg/kg subcutaneously for 7 days) attenuated the rise in endothelaemia and adrenal corticosterone measured at 24 h following IMO. Plasma levels of ACTH and proopiomelanocortin gene expression in the anterior pituitary were not affected by pentoxifylline treatment. The present results indicate that IMO stress in female rats induces a biphasic rise in endothelaemia early at the time of stress exposure and than 24 h thereafter. Based on these data and our previous study we can conclude that intensive stress has a negative influence on endothelial cells in both sexes and no gender differences seem to be present in the protective action of pentoxifylline.     

重复制动应激对雌性大鼠下丘脑-垂体-卵巢轴的影响

目的: 探索重复制动应激对雌性大鼠下丘脑-垂体-卵巢轴的影响。方法: 40只SD雌鼠随机分为两组(n=20),对照组和实验组,一组正常饲养,一组采取递增负荷束缚应激,每天置于束缚器内制动应激一次(从上午9:00开始),第1日制动2 h,以后采用递增负荷,每日增加0.5 h,持续两周,通过检测体重、脏器系数、动情周期、性激素、病理和相关基因的表达探索其对下丘脑-垂体-卵巢轴的危害。结果: 重复制动应激使雌性大鼠体重下降、动情周期延长,卵巢和子宫的脏器系数和形态发生改变,利用qPCR技术对其相关基因检测,发现下丘脑促性腺激素释放激素、垂体促性腺激素释放激素受体、促卵泡生成素和促黄体生成素mRNA的表达显著下降,卵巢促卵泡生成素和黄体生成素受体 mRNA的表达显著上升,卵巢和子宫雌激素受体mRNA的表达显著下降。结论: 重复制动应激可能通过干扰下丘脑-垂体-卵巢轴的内分泌调节作用,使动情周期紊乱,从而损伤雌性动物的性腺和生殖内分泌功能。     

Effect of 2-deoxy-D-glucose on hypothalamic-pituitary-thyroid axis in rats

The intraperitoneal administration to rats of 500 mg/kg body weight of 2-deoxy-D-glucose, an analog of glucose which produces intracellular glucopenia with rise in extracellular fluid glucose concentration, is followed by a significant though transient reduction of hypothalamic TRH content, observed at 15 and 25 minutes after drug administration. A subsequent increase in serum thyrotropin followed by that of triiodothyronine concentration was also observed. These findings indicate that the neuroglucopenia induced by 2-deoxy-D-glucose may play a role in the regulating the hypothalamic-pituitary-thyroid axis.     

Disorders of sexual development and associated changes in the pituitary-gonadal axis in dogs

Normal sexual differentiation depends on completion of chromosomal sex determination, gonadal differentiation, and development of the phenotypic sex. An irregularity in any of these three steps can lead to a disorder in sexual development (DSD). We examined nine dogs with DSD by abdominal ultrasonography, laparotomy, histologic examination of the gonads, and reproductive tract, cytogenetic analysis, and mRNA expression of the SRY gene. We also determined the plasma concentrations of luteinizing hormone (LH), estradiol-17β, and testosterone before and after administration of gonadotropin-releasing hormone (GnRH) and compared these results with those obtained in anestrous bitches and male control dogs. The gonads of three dogs with DSD contained both testicular and ovarian tissue, while in the other six only testicular tissue was found. Each of the dogs had a uterus. Based on gynecologic examination, cytogenetic analysis, and the histology of the gonads, seven of the nine dogs appeared to be XX sex reversals. Three of these were XX true hermaphrodites and four were XX males; the other two dogs had incomplete XY gonadal dysgenesis. All seven XX sex-reversed dogs were found to be negative for the SRY gene by polymerase chain reaction. The basal plasma luteinizing hormone (LH) concentration was significantly higher in dogs with DSD than in anestrous bitches but not significantly different from that in male dogs. The basal plasma LH concentration increased significantly after GnRH administration in all dogs with DSD. The basal plasma estradiol concentration was significantly higher in dogs with DSD than in anestrous bitches but not significantly different from that in male dogs. The basal plasma testosterone concentration was lower in dogs with DSD than in male dogs. In all dogs with DSD both the basal and GnRH-induced plasma testosterone concentrations were above the upper limit of their respective ranges in the anestrous bitches. In conclusion, the secretion of LH and estradiol in these dogs with DSD, all of which had testicular tissue in their gonads, was similar to that in male control dogs. These results indicate that the basal and/or GnRH-stimulated plasma testosterone concentration might be used to detect the presence of testicular tissue in dogs with DSD.     

Reproductive axis response to repeated lipopolysaccharide administration in peripubertal female rats

Immune system disorders are often accompanied by alterations in the reproductive axis. Several reports have shown that administration of bacterial lipopolysaccharide (LPS) has central inflammatory effects and activates cytokine release in the hypothalamus where the luteinizing hormone releasing hormone (Gn-RH) neurons are located. The present study was designed to investigate the effect of repeated LPS administration on the neuroendocrine mechanisms of control of the reproductive axis in peripubertal female rats (30-day-old rats). With this aim, LPS (50 μg/kg weight) was administered to the animals during 25, 27 and 29 days of age and sacrificed on 30 day of life. Gn-RH, γ?amino butyric acid (GABA) and glutamic acid (GLU), two amino acids involved in the regulation of Gn-RH secretion, hypothalamic content were measured. LH and estradiol serum levels were also determined and the day of vaginal opening examined. The results showed a significant increase in Gn-RH and GLU content (p?<?0.0001), shared by a reduction of GABA one (p?<?0.0001). LH and estradiol serum levels were decreased (p?<?0.01, p?<?0.001) and delay in the day of vaginal opening was also observed in treated animals. Present results show that repeated LPS administration impaired reproductive function, modifying the neuroendocrine mechanisms of control of the axis in peripubertal female rats.     

Effects of an LHRH agonist on endocrine function, LHRH receptor and LH/hCG receptor in the pituitary-gonadal axis of male rats

The effects of an LHRH agonist (LHRHa), [D-Ser (tBu)]6 des-Gly-NH210) ethylamide, on endocrine function and the LHRH and LH/hCG receptors in the pituitary-gonadal axis were examined. The LHRHa was injected at 100 ng/100 g body weight into male rats once a day for 4 weeks and its effects were observed until 2 weeks after the end of treatment. Due to LHRHa treatment, the plasma LH concentration began to increase on day 3, reached a peak on day 7, and then decreased, although it remained above the control level during the treatment. The pituitary LH content decreased on day 1, reached a minimum (about 40% of the control) between days 3 and 7, and then was maintained at 60% of the control level until week 4. In contrast, the pituitary LHRH receptor concentration increased only on day 3, and the association constant (Ka) remained unchanged during the observation period. The testis weight and plasma testosterone concentration began to decrease on day 3, reached the minimum on day 7 and remained at this level until week 4, and their levels were not completely restored to normal 2 weeks after cessation of treatment. The testicular LH/hCG receptor concentration was decreased on day 1, and markedly decreased to 10-15% of the control value between day 7 and week 4, but the Ka value was slightly increased during the treatment. However, these values had completely recovered 2 weeks after the cessation of treatment. The testicular LHRH receptor concentration increased between days 1 and 7, returned to the control level in weeks 2 and 4, and then decreased 2 weeks after cessation of treatment. Its Ka value was reduced in weeks 2 and 4. These data suggest that the inhibitory effect of LHRHa on the gonad in male rats is not due to reduced pituitary LH release, but to changes in the number and Ka values of gonadal receptors for LH/hCG and LHRH.     

Inhibition of pituitary-gonadal axis in mice by long-term administration of D-Trp-6-LHRH microcapsules

Female mice were injected, every 30 days for 5 months, with a long-acting formulation of microcapsules liberating 2.5 micrograms D-Trp-6-LHRH/day. The control group was injected with vehicle only. At 30 days after the last injection mice were killed, ovaries, uteri and adrenals were weighed and fixed in formalin for histological studies. LH and oestradiol concentrations were measured by RIA. In the D-Trp-6-LHRH-treated group, the weights of the ovaries and uterus (P less than 0.01 and P less than 0.05, respectively), and LH and oestradiol values (P less than 0.02 and P less than 0.01, respectively) were reduced compared to controls. Histologically, the ovaries contained a large number of degenerated, atretic follicles, and corpora lutea had almost completely disappeared. These results indicate, contrary to the prevailing opinion, that mice are sensitive to inhibitory effects of LHRH agonists and that a suppression of the pituitary-gonadal axis can be obtained with long-term administration of D-Trp-6-LHRH microcapsules.     

Effects of early experience on female behavioural and reproductive development in rhesus macaques

A growing body of research on humans suggests that exposure to a stressful family environment or father absence from home during childhood is associated with early female puberty and greater interest in infants among adolescent girls. This effect may be mediated by early exposure to harsh and inconsistent maternal care, but the mechanisms by which maternal care affects female reproductive maturation are not known. The present study reports sex differences in interest in infants among juvenile rhesus macaques similar to those observed in human adolescents. Furthermore, juvenile females that were exposed to harsh and inconsistent maternal care in infancy showed higher interest in infants than controls. Evidence from cross-fostered females indicated that these effects resulted from early experience and not genetic inheritance from the mother. There were no significant differences in female age at first conception in relation to the quality of maternal care received during infancy. Macaque females exposed to harsh and inconsistent maternal care in infancy tended to have higher cortisol responses to stress and to corticotropin-releasing hormone than controls in the first three years of life. Furthermore, females with higher cortisol responses to stress exhibited higher interest in infants. These findings suggest that some of the effects of early parental care on female reproductive maturation may be mediated by developmental changes in the activity of the hypothalamic-pituitary-adrenal axis.     

Effect of neonatal androgenization on the septal neurons of the brain in female rats in early postnatal ontogeny

Newborn female rats were androgenized, and the reaction of neurons of brain septum on excessive quantity of exogenous androgens, introduced during so-called "crucial" period of formation of centers of gonadotropic regulation of sexual cycles, has been studied in 3, 5, 7, 10, 20, 30, and 60 days old animals. Morphometry of brain septum cell nuclei revealed that most neuron nuclei shrink after androgenization. Monoamine content was significantly increased in septum nuclei of experimental animals. Neonatal androgenization led to the increased capacity of septal complex neurons to bind 3H-estradiol and to the decreased 3H-testosterone binding. The data obtained suggest that the brain septum neurons of female rats depend on sex steroids, particularly during "crucial" period of development.     

Functional and morphological changes in the hypothalamo-pituitary-gonadal axis of aged female rats.

Age-related functional and morphological alterations in the hypothalamo-pituitary-gonadal axis were investigated in old recurrently pseudopregnant (RPP) female rats, and these alterations were compared with those in young diestrous rats. LHRH in the median eminence (ME) and mediobasal hypothalamus (MBH) as well as plasma FSH, LH, and progesterone were measured by RIA. LHRH in the lateral ME (LME) and pituitary FSH and LH were evaluated by morphometry and densitometrical immunocytochemistry. Furthermore, by light microscopy, we classified and counted the number of ovarian follicles and corpora lutea. LHRH concentrations in the ME and MBH were similar in old and young rats, whereas in old rats, plasma FSH was markedly increased, LH was moderately increased, and plasma progesterone was unchanged. The number and the total area and immunoreactivity of LHRH-labeled axon cross sections in the LME were reduced in old rats. The number of nucleated FSH-labeled cells and total FSH area and immunoreactivity were almost twice in old compared with young animals. The measurements of LH-labeled cells were not different between the two groups. In old rats, the numbers of ovarian follicles and corpora lutea were reduced and that of atretic follicles increased. In conclusion, age-related morphological impairments of LHRH axons associated with an increased number of FSH gonadotropes and higher plasma FSH in our old RPP rats suggest hypothalamic and pituitary disturbances, which may largely contribute to the complex hormonal disarrangement responsible for the decline of reproductive functions in old female rats.     

Effects of acute alcohol intoxication on pituitary-gonadal axis hormones, pituitary-adrenal axis hormones, beta-endorphin and prolactin in human adolescents of both sexes

Teenage drinking continues to be a major problem in industrialized countries, where almost 35% of alcohol drinkers are under 16 years old. In the present paper we studied the effects of acute alcohol intoxication (AAI) on the pituitary-gonadal (PG) axis hormones, and the possible contribution of pituitary-adrenal (PA) axis hormones, beta-endorphin (BEND), and prolactin (PRL) to the alcohol-induced dysfunction of PG axis hormones. Blood samples were drawn from adolescents that arrived at the emergency department with evident behavioral symptoms of drunkenness (AAI) or with nil consumption of alcohol (controls [C]). Our results demonstrated that AAI produces in adolescents a high increase in plasma PRL, ACTH, and cortisol (F), and a contradictory behavior of testosterone (T) according to gender: plasma T was increased in females and decreased in males. ACTH and PRL correlated positively with F, dehydroepiandrosterone-sulphate (DHEAS) and T in females, which suggests that PRL and ACTH could synergistically stimulate adrenal androgen production. In contrast, the decrease in T and increase in BEND in males suggests that AAI could have an inhibitory effect on testicular T, perhaps mediated by BEND. The hormones studied are involved in the development of secondary sexual characteristics and the growth axis during adolescence. The deleterious effects of alcohol abuse should be made known to adolescents and the appropriate authorities.     

The pituitary-gonadal axis before puberty: Effects of various estrogenic steroids in the ovariectomized rat

A series of experiments were conducted to determine the effects of several estrogens upon FSH and LH secretion in immature ovariectomized rats. Groups of animals were castrated at 26 days of age and treated for 5 days post-operatively with various dosages of one of the following steroids: estrad1ol-17β(E₂), estradiol benzoate (EB), estrone (E₁), equilenin (EQ), 17α-ethinyl estradiol (EE), or mestranol (ME). Uterine weights were recorded and blood taken for radioimmunoassay.Estradiol was able to suppress both FSH and LH within a “physiologic dosage range” (PDR), wherein both gonadotropins were suppressed to intact control levels by a dose which did not stimulate uterine weight higher than intact control weight. EB and ME suppressed LH but not FSH at the PDR; the other steroids suppressed at higher than PDR doses. LH was preferentially suppressed, as compared to FSH, by all 6 steroids. By biological potency the order of activity was E₂ = EE ? EB ? ME ? E₁ ? EQ. For relative ability to suppress FSH (compared to LH), the order was E₁ or E₂, ? ME ?EB ? EE ? EQ. At higher doses (near maximum uterine stimulation), e₁, E₂ and EE produced higher FSH and LH than suppressed levels seen at lower doses; a pharmacologie dosage of E₂ caused re-suppression of both gonadotropins.Results indicate that a feedback system is present before puberty and this system is sensitive to very low levels of estrogens. Likewise, there is a potential for positive feedback present for higher estrogen levels, and a complete suppression occurs at pharmacologie levels. There appears to be a significant discrepancy between the biologic activity (by uterine weight) of estrogens and their ability to affect gonadotropin