Alterations in the onset of ovulatory failure and gonadotropin secretion following steroid administration to lightly androgenized female rats

The present studies were designed to characterize the gonadotropin response to exogenous steroids in neonatally androgenized female rats in various states of reproductive decline. Female rats were androgenized by the administration of a single injection of testosterone propionate (TP) (10 or 100 micrograms) at 5 days of age. Control rats received sesame oil. Treatment with 100 micrograms TP resulted in persistent vaginal estrus (PVE) from the onset of vaginal introitus. Treatment with 10 micrograms TP resulted in a period of regular estrous cyclicity followed by PVE. In the first experiment, all animals were ovariectomized between the ages of 60-85 days and the gonadotropin response to exogenously administered estradiol benzoate (EB) (10 micrograms/100 g BW) and progesterone (P) (2 mg/animal) was determined. When testing began 3 days following ovariectomy, control females exhibited significant (P less than 0.01) afternoon elevations of luteinizing hormone (LH) and follicle-stimulating hormone (FSH) following EB, which were further amplified following P. When ovariectomy occurred prior to the onset of PVE (PRE PVE), lightly androgenized females (10 micrograms TP) showed no significant afternoon gonadotropin increase following EB. Following P, phasic LH secretion was present but significantly (P less than 0.01) decreased in amplitude and delayed in onset versus that of control females. When ovariectomy occurred 3 to 4 wk following the onset of PVE, lightly androgenized females (PVE group) as well as fully androgenized females (FAS) (100 micrograms TP) showed no gonadotropin response to steroid priming.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effect of steroid aromatase inhibitors on the catecholamine content of the hypothalamus of neonatally androgenized rats

Administration of 50 micrograms of testosterone propionate to newborn female rats on the 5th day of life provoked a reliable increase in noradrenaline and dopamine concentrations in the hypothalamus of 10-day-old rats. Neonatal administration of aromatase inhibitors on the 5th and 7th days of life prevented testosterone-induced increase in catecholamine concentrations. The data obtained prove the integration of the processes of testosterone aromatization and catecholamine accumulation in androgen-dependent brain differentiation.     

Hypothalamic-pituitary somatotropic function in prepubertal hypothyroid rats: effect of growth hormone replacement therapy.

The effect of thyroid hormone deficiency and growth hormone (GH) treatment on hypothalamic GH-releasing hormone (GHRH)/somatostatin (SS) concentrations, GHRH/SS mRNA levels, and plasma GH and somatomedin-C (IGF-I) concentrations were studied in 28- and 35-day-old rats made hypothyroid by giving dams propylthiouracil in the drinking water since the day of parturition. Hypothyroid rats, at both 28 and 35 days of life, had decreased hypothalamic GHRH content and increased GHRH mRNA levels, unaltered SS content and SS mRNA levels, and reduced plasma GH and IGF-I concentrations. Treatment of hypothyroid rats with GH for 14 days completely restored hypothalamic GHRH content and reversed the increase in GHRH mRNA, but did not alter plasma IGF-I concentrations. These data indicate that, in hypothyroid rats, the changes in hypothalamic GHRH content and gene expression are due to the GH deficiency ensuing from the hypothyroid state. Failure of the GH treatment to increase plasma IGF-I indicates that the feedback regulation on GHRH neurons is operated by circulating GH and/or perhaps tissue but not plasma IGF-I concentrations. Presence of low plasma IGF-I concentrations would be directly related to thyroid hormone deficiency.     

Brain monoamines and sexual behavior in Japanese quail: Effects of castration and steroid replacement therapy

In the male Japanese quail, testosterone is required for the activation of sexual behavior. This steroid dependent process may rely heavily on mediation via monoaminergic neurons. These experiments were conducted to study the relationship between reproductive state (hormonal and behavioral components) and levels of monoamines in selected areas of the brain in Japanese quail. In Experiment 1, monoamine levels in a number of brain areas were compared in castrates, testosterone-implanted castrates, and intact males. Monoamine levels were comparable to those previously measured in Japanese quail, and there were no significant differences due to treatment. Plasma luteinizing hormone (LH) levels and recovery of cloacal gland area in implanted castrates confirmed the afficacy of treatments. In Experiment 2, the disappearance of dopamine (DA) and norepinephrine (NE) following administration of a-methyl-para-tyrosine (aMPT) was used as an indicator of turnover rate. Male and female quail were gonadectomized at 3 weeks of age. At the age of five weeks, some gonadectomized males and females were given implants containing testosterone. Only intact males and testosterone-implanted castrated males showed reproductive behavior. Plasma gonadotropin levels were elevated in gonadectomized birds and reduced in steroid-implanted gonadectomized birds. The aMPT treatment significantly reduced the levels of DA and NE in the telecephalon and the level of DA in the hypothalamus. After aMPT treatment, the disappearance of NE in the telecephalon and of DA in the hypothalamus were significantly different according to the sex or treatment of the birds or both. Significant interactions between these two factors were observed. Disappearance rate of NE in the telecephalon was decreased by castration of males and increased by ovariectomy of females. Both effects were counteracted by testosterone. Reverse effects were observed for DA disappearance in the hypothalamus (increase with castration in males and decrease with ovariectomy in females). These results give evidence for altered aminergic function in specific areas of the brain relative to altered reproductive state.     

Leptin levels in menopause: effect of estrogen replacement therapy

To evaluate the effect of menopause and estrogen replacement therapy on leptin levels, 17 white postmenopausal women were recruited for the study. After an overnight fasting, blood samples were collected for LH, FSH, estradiol, testosterone, androstenedione, DHEA sulfate, insulin and leptin assays. Body mass index (BMI) and the waist-to-hip ratio were also evaluated. Patients were reanalyzed after a 12-week administration of transdermal estrogen patches delivering 50 microg 17beta-estradiol. The results were compared to those obtained from a group of 11 female volunteers in reproductive age, in whom basal blood was sampled during the early follicular phase of their cycle. Patients were divided into lean and obese according to their BMI. Obese postmenopausal women showed lower leptin levels when compared to premenopausal counterparts (25.1 +/- 5.9 vs. 37 +/- 11.3; p < 0.05), whereas no significant differences were found between the lean groups (14.5 +/- 3.8 vs. 14.4 +/- 4.9). Estrogen administration did not significantly change serum leptin concentrations in hypoestrogenized women (obese: 25.1 +/- 5.9 vs. 28. 6 +/- 9.2; lean: 14.4 +/- 4.9 vs. 17.6 +/- 7.2). A positive linear correlation was found between leptin plasma levels and BMI only in obese patients (r = 0.58; p < 0.01) both before and after estrogen treatment. Menopause is characterized by a decreased expression of the obese gene, even if estrogens do not seem to represent a main causal factor.     

Activities of glycolytic enzymes in some brain areas of thyroidectomized rats and their response to replacement therapy

Glycolytic metabolism has been assessed by studying a set of key enzymes, in anterior cortex, amygdala, hypothalamus septum and hippocampus, in thyroidectomized rats. The reversibility of the changes induced by the thyroidectomy has been assessed by replacement therapy. In thyroidectomized rats the hexokinase activity was significantly decreased in anterior cortex and hypothalamus. The increase in phosphofructokinase and pyruvate kinase activity was probably due to an increase in cellular energy requirements. Hexokinase activity was best restored by treatment with L-thyroxine (T4) or T4+ propylthiouracil (PTU). The low response of pyruvate kinase activity in all treated animals could suggest that this metabolic step is the least reversible.     

Spontaneous running activity in male rats: effect of age

Alterations in the intensity and pattern of spontaneous running activity as rats increase in age from 7 wk to 1 yr was studied in male rats placed in exercise wheel cages. Daily running records were obtained on 27 rats for periods up to 12 mo, and 24-h activity recordings were made of selected runners to study the variation in activity during the day. The data indicate that for rats running over 2,940 revolutions (or 2 miles/day), the maximum intensity of running attained can be divided into a group of high achievers (approximately 8 miles/day) and moderate achievers (averaging 4.5 miles/day). For both groups, spontaneous running activity reached maximal rates after 4-5 wk. This maximal rate was sustained for 7-8 wk, then fell to levels approximately 60% of maximum for 4-5 mo, and then fell again to levels approximately 25% of maximum from 8 to 12 mo of age. The hourly pattern of running activity during the day was defined in rats of increasing age, who averaged 13,280, 6,662, 3,874, and 1,755 rev/day, corresponding to 9.0, 4.5, 2.6, and 1.2 miles/day, respectively. The overall patterns at each level indicated that the major running period occurred between 6:00 P.M. and 6:00 A.M., the greater activity of younger rats was paralleled by faster speeds and longer duration at each hour of the day, and the peak running activity for each group generally occurred between 7:00 and 9:00 P.M. In summary, there is a progressive loss in speed and duration of spontaneous running activity as male rats increase in age, with intensity of exercise falling below 2 miles/day after 7-8 mo of age.     

Luteinizing hormone response to N6, O2'-dibutyryl adenosine 3',5'-cyclic monophosphate in aged, young and androgenized rats: estradiol effect in vitro

The studies in this report were designed to investigate whether the loss of pituitary luteinizing hormone (LH) responses to N6,O2'-dibutyryl adenosine 3',5'-cyclic monophosphate (DBcAMP) in aged noncycling rats was the result of age, endocrine status associated with diestrus, or noncyclic low estrogen status. Pituitary monolayer cultures were prepared from female Fischer 344 rats. Aged (18-month-old) persistent diestrous (PD) rats, young diestrous (D) rats, or noncycling neonatally androgenized-constant estrous (AN-CE) rats were used. Enzymatically dispersed cells were maintained in the same batch of medium supplemented with dextran-coated charcoal adsorbed serums. Total LH contents were 1.75 +/- 0.04, 1.15 +/- 0.03, and 1.71 +/- 0.02 micrograms LH/dish in Day 5 cultures prepared from aged PD, young D, and AN-CE rats, respectively. Incubations with 5 mM DBcAMP for 4 h significantly (P less than 0.05) stimulated LH release in cultures prepared from young D and AN-CE rats but inhibited LH release in cultures prepared from aged PD rats even though a 4-h incubation with 10 nM LH releasing hormone (LHRH) stimulated LH release similarly in cultures of all three types of cells. The loss of DBcAMP-induced LH release in cultures prepared from aged PD rats was reversed by 17 beta-estradiol (E2). This treatment also reduced the basal LH release and increased the cellular LH content. These results indicate that the loss of DBcAMP-induced LH response in the aged rat is not an irreversible aging phenomenon but appears to be associated with the chronically low E2 status of aged PD rats but not young cycling D or noncycling AN-CE rats.     

Testicular binding of 125I-HCG in developing estrogenized and androgenized rats.

The present work describes changes seen in the binding of 125I-HCG by testis homogenates of rats injected at the age of three days with 400 microgram estradiol-17beta-dipropionate, or 250 microgram testosterone propionate. Estrogenized and androgenized male rats showed a marked decrease of gonadotropin binding in testis at the 30th, 45th and 60th postnatal day. These results show that delayed sexual maturation of male rats treated with estrogen and androgen in the neonatal period is also related to pubertal decrease of testicular gonadotropin receptors.     

The use of embryo transfer and steroid hormone replacement therapy in the study of prenatal mortality

Embryo transfer and steroid hormone replacement therapy have been used to advantage in studies on prenatal mortality, particularly in sheep.The form, timing and magnitude of loss, have been well documented and steroid hormone replacement therapy coupled with embryo transfer has clearly shown the involvement of the ovarian steroid hormones secreted before, during and after estrus in survival and development of embryos.However, little is known of the manner in which the steroid hormones exert their effects on the environment of pre-implantation embryos; a stage when most mortality occurs.