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Effect of gonadal steroids on hypothalamus and anterior pituitary endo-oligopeptidase B (proline-endopeptidase) activity in castrated female rats.

In the present study we investigated the possible participation of endo-oligopeptidase B (poline-endopeptidase) in the control of gonadotrophin secretion through the control of LH-RH inactivation. This enzyme selectively hydrolyzes the Pro9-Gly10-NH2 peptide bond of LH-RH, thereby inactivating this substance. The enzyme activity was evaluated using a specific colorimetric substrate, i.e., Z-Gly-Pro-SM. Female adult Wistar rats were submitted to castration, experimental situations that are known to produce changes in gonadotrophin secretion. Hypothalamic and pituitary endo-oligopeptidase B activity was shown to be present predominantly in the soluble fraction of the enzyme preparations. The results also indicated that endo-oligopeptidase B activity adult female rat pituitary decreased after castration and increased after administration of estradiol and progesterone to castrated animals. The present results lead us to suggest that anterior pituitary endo-oligopeptidase B may be related to the control gonadotrophin secretion in female rats.

Animals↗

Mediation by gonadal steroids of plasma beta-endorphin and LH in castrated female and male rats.

Immunoreactive beta-endorphin (IR-BE) was significantly decreased and luteinizing hormone (LH) significantly increased in female rats castrated for four weeks. Forty eight hours after a single injection of estradiol benzoate (EB), IR-BE levels increased, and LH levels were reduced. On the afternoon following the administration of a second injection of EB given six hours earlier, IR-BE levels were reduced below control values, whereas LH levels were significantly elevated. There was no change in IR-BE levels during the remainder of that afternoon whereas LH levels decreased over time. Similar to female rats, IR-BE was diminished and LH increased in castrated male rats. IR-BE was increased significantly above those values observed in intact animals 24 hr after a single injection of TP and returned to control levels by 48 hr after administration of TP. Injection of TP reduced LH to levels observed prior to castration. These findings suggest that gonadal steroids exert a feedback on the release of IR-BE from the pituitary of female and male rats opposite to their feedback effect on the release of pituitary gonadotropins.

Animals↗

Seasonal levels of minerals, enzymes, nutrients and metabolic products in plasma of intact and castrated adult male white-tailed deer (Odocoileus virginianus).

1. Alkaline phosphatase (AP), cholesterol, creatinine, uric acid, total protein, albumin, bilirubin, urea nitrogen, calcium, glucose, lactic dehydrogenase (LDH) and serum glutamic-oxalacetic transaminase (SGOT) were measured in the plasma of three intact and three castrated male deer. 2. A statistically significant seasonal cycle of AP, cholesterol, creatinine and uric acid was found in intact but not in castrated animals. 3. Monthly levels of total protein, albumin, urea nitrogen, bilirubin and calcium were significantly higher in castrated deer. 4. On the other hand, monthly levels of LDH and SGOT were higher in intact animals.

Alkaline Phosphatase↗

Effect of several androgens, cyproterone acetate or estrogen-progesterone on the prolactin-releasing activity of arginine vasotocin in castrated male rats.

Intravenous (iv) administration of 5 microgram of arginine vasotocin (AVT) into urethane-anesthetized, castrated male rats had no effect on plasma prolactin titers as compared to the rise in prolactin levels observed in intact AVT-treated rats. However, when castrated rats were first treated for two days with 2.5 mg/day of testosterone propionate and then challenged with a 5-microgram dose of AVT, the prolactin surge values obtained were comparable to those seen in intact AVT-treated rats. Conversely, treatment of intact rats for two days with 25 mg/day of the anti-androgen, cyproterone acetate, blocked the prolactin-releasing activity of AVT. In a separate experiment, treatment of castrated rats for two days with 2.5 mg/day of testosterone, androsterone or 50 microgram of estradiol benzoate and 25 mg progesterone, completely restored the prolactin-releasing activity of AVT. Similar treatment with 2.5 mg/day of androstenedione or dihydrotestosterone was without effect in restoring this response to AVT. It is concluded that the presence of gonadal steroids is essential to the action of AVT in provoking the release of prolactin in urethane-anesthetized male rats.

Androgens↗

Differential effects of castration and testosterone replacement on basal and LHRH-stimulated cAMP and cGMP accumulation and on gonadotropin release from the pituitary of the male rat.

Castration of male rats decreased cAMP levels, and increased cGMP levels and gonadotropin release from anterior pituitaries incubated in vitro. Testosterone (T) replacement via silastic tubes filled with the steroid increased cAMP and decreased cGMP levels and gonadotropin release. Incubation of hemipituitaries from intact males with luteinizing hormone releasing hormone (LHRH, 5 nM for 2 h) resulted in increased cAMP and cGMP accumulation and gonadotropin release. Castration abolished LHRH-induced cAMP accumulation, but increased the effect of LHRH on cGMP accumulation and gonadotropin release. Testosterone replacement restored cAMP stimulation by LHRH but decreased LHRH-induced elevation of cGMP levels and gonadotropin release. These data illustrate parallel increases by castration of LHRH-induced cGMP accumulation and of gonadotropin release. Furthermore, these two parameters are influenced in the opposite direction by replacement therapy. These results support the concept of a role for cGMP in LHRH action as well as providing evidence of a link between the feedback action of T and cGMP in the pituitary gland.

Animals↗

Effects of ethanol treatment and castration on liver alcohol dehydrogenase activity.

Induction of alcohol dehydrogenase (ADH) activity by chronic ethanol treatment and castration has previously been reported to occur in Sprague-Dawley rats. In the present study, no induction was found following chronic ethanol treatment and only a low level of induction was found with castration. However the activity of ADH was high in control animals compared with those used in other studies. The activity of ADH in control animals was not decreased by testosterone administration, which has been shown to reverse induction of the enzyme produced by chronic ethanol treatment or castration in other studies. It is concluded that the male Sprague-Dawley rat is not necessarily a suitable animal for the study of ADH induction by chronic ethanol treatment and that further unknown factors must be identified before the regulation of ADH activity in vivo is fully understood.

Alcohol Dehydrogenase↗

Genotype and retention of the ejaculatory reflex in castrated male mice.

Genotype proves to be an important factor affecting the retention of sexual responsiveness following castration. Male F1 hybrids between C57BL/6 females and DBA/2 males (B6D2F1) show very prolonged retention. Experiment 1 used a diallelic design using C57BL/6, DBA/2 and BALB/c strains and showed that heterosis per se is not an adequate explanation of the superior retention of the B6D2F1 genotype. Experiment 2 showed that neonatal injections of 100 mug TP to B6D2F1 male mice did not result in better retention of the ejaculatory reflex after castration in adulthood. B6D2F1 males from both experiments who exhibited the ejaculatory reflex for many months after castration went through a 'difficult period' in the first 10 to 12 postcastration weeks. During this time, ejaculation latency increased fourfold, and then returned to the original level. On the other hand, the proportion of males reaching the ejaculatory threshold during this period first declined then increased.

Animals↗

Castration rapidly decreases hypothalamic gamma-aminobutyric acidergic neuronal activity in both male and female rats.

The postcastration LH response is greater and somewhat more rapid in male than female rats. We have previously demonstrated that hypothalamic gamma-aminobutyric acid (GABA)ergic neuronal activity decreases following gonadectomy in male rats. To investigate whether these same hypothalamic GABA neurons decrease their activity postcastration in female rats, and whether more rapid and or greater postcastration decreases occur in male rats, we determined the timing and magnitude of the postcastration decreases in GABA turnover which are associated with the sexually dimorphic postcastration LH response. Adult male and 4-day cycling female rats were castrated between 0800 and 1000 h (females ovariectomized on diestrus day 1). Serum LH levels increased significantly by 12 h postcastration in both males and females with the magnitude of the increases being 6.2-fold in males and 2.8-fold in females. GABA turnover was determined in 16 microdissected brain structures by the GABA transaminase inhibition method at 0 h (sham-operated controls), 6 h, 12 h and 1, 2, 4 and 6 days postcastration. In male rats, in the diagonal band of Broca at the level of the organum vasculosum of the lamina terminalis [DBB(ovlt)], the rate of GABA turnover decreased significantly already by 6 h postcastration compared with the 0 h controls, and remained suppressed through 6 days. This rapid down regulation of DBB(ovlt) GABAergic neurons also occurred in female rats, however, the duration of the decrease was not as prolonged as in male rats. Similar changes occurred in the tuberoinfundibular GABAergic (TIGA) neurons projecting to the median eminence in both males and females. Down regulation of these GABAergic neurons precedes or is coincident with increased postcastration LH secretion in both sexes, and the duration of the decreases is consistent with the less robust postcastration LH response in female rats. In addition, the rate of GABA turnover decreased after castration in the interstitial (bed) nucleus of the stria terminalis, ventral aspect (INSTv), the medial preoptic nucleus, dorsomedial aspect (MPNdm) and the ventromedial nucleus, ventrolateral aspect (VMNvl) in male rats, and in the INSTv and VMNvl of female rats, while there was no effect of castration in other hypothalamic regions or control structures. The result in the female VMNvl is consistent with reports that GABA facilitates lordosis behavior in this hypothalamic structure. These findings are consistent with the hypothesis that discrete hypothalamic populations of sex steroid-sensitive GABAergic neurons mediate the postcastration LH responses in both male and female rats, and may underlie other sexually dimorphic adult phenotypes such as sex behavior.

Animals↗

Functional effects of castration on alpha1-adrenoceptors in rat vas deferens.

The effects of castration on alpha1-adrenoceptors in rat vas deferens were investigated by determining the actions of selective antagonists against the contractions induced by noradrenaline. The results obtained in vas deferens from control rats suggest participation of alpha1A-adrenoceptors as judged by the pA2 values for prazosin (9.6), benoxathian (9.5), 2(2,6-dimethoxyphenoxyethyl) amino-methyl-1,4-benzodioxone hydrochloride) (WB 4101) (9.6), phentolamine (8.4), 8-[2-[4-(2-methoxyphenyl)-1-piperazinyl]ethyl]-8-azaspiro[4.5] decane-7,9-dionedihydrochloride (BMY 7378) (6.7) and by the insensitivity to chloroethylclonidine (100 microM, 45 min). In vas deferens from castrated rats, WB 4101 and spiperone showed slopes lower than 1.0 in the Schild plots, suggesting participation of multiple receptors. In these organs, noradrenaline contractions were partially inhibited by chloroethylclonidine (100 microM, 45 min), indicating participation of alpha1B-adrenoceptors. After chloroethylclonidine treatment, WB 4101 showed a slope not different from 1.0 in the Schild plot, resulting in a pA2 of 9.4, which indicates an interaction with alpha1A-adrenoceptors. It is suggested that castration modifies the functional alpha1-adrenoceptors subtypes in rat vas deferens.

Adrenergic alpha-1 Receptor Antagonists↗

Investigations into the effect of castration on the blood glucose profile of male and female rats treated with glibenclamide (HB 319).

In SPF-bred male and female Sprague-Dawley rats the effect of castration on the hypoglycaemic action of a single dose of glibenclamide (HB 419) was investigated. The results of castration showed no unambiguous influence on the normal blood glucose values. The effect of this sulphonyl urea, however, was considerably stimulated by castration and was prolonged in male animals. Obviously the absence of testosterone has an inhibitory effect on the activity of liver ribosomes thus causing a delay in the metabolism. Furthermore the metabolic rate depends on the testosterone value in the organism as evidenced by the differences between male and female animals. The results were ascertained statistically.

Animals↗

Tachykinins in the pineal gland: effect of castration and ganglionectomy.

In this investigation, the presence of NKA-immunoreactive substances was determined in pineal glands from intact, castrated and castrated, testosterone-treated male rats. The effect of environmental light, melatonin treatment and superior cervical ganglionectomy on pineal NKA-immunoreactive substances was also investigated. The results obtained show that NKA is present in measurable amounts in the rat pineal, and NPK is probably also present, Orchidectomy was followed by an increase in the content of NKA-immunoreactive substances in the pineal gland. The replacement treatment with testosterone propionate in castrated rats blocked this effect. NKA-immunoreactive substances were not significantly different quantitatively in pineals from rats killed under light or under darkness. The removal of the superior cervical ganglia was followed by a significant increase in the NKA-immunoreactive substance content in the pineal gland of male rats. These results indicate that NKA and other tachykinins are present in the pineal gland of the male rat, and they seem to be regulated by gonadal hormones and the innervation originated from the superior cervical ganglia.

Animals↗

Cytochrome oxidase activity in the preoptic area correlates with differences in sexual behavior of intact and castrated male leopard geckos (Eublepharis macularius).

Although the utility of analyzing behavioral experience effects on neural cytochrome oxidase (CO) activity is well recognized, the behavioral correlates of endogenous differences in CO activity have rarely been explored. In male leopard geckos (Eublepharis macularius), the incubation temperature experienced during embryogenesis (IncT) and age affect CO activity in the preoptic area (POA), an area that modulates copulatory behavior. In this study, the authors assessed whether differences in POA CO activity correlate with differences in sexual behavior in intact and castrated geckos. Males with IncT- and age-dependent increases in POA CO activity mounted females with shorter latencies while intact and after castration and ejaculated more frequently after castration. The authors discuss the predictive value of CO activity and propose similar parallels in other species.

Age Factors↗

Deoxyribonucleic acid and polyamine synthesis in rat ventral prostrate. Effects of age of the intact rat and androgen stimulation of the castrated rat with testosterone, 5 alpha-dihydrotestosterone and 5 alpha-androstane-3 beta, 17 beta-diol.

The relationship between polyamine synthesis, growth and secretion in vivo was examined in ventral prostates from: (a) intact rats aged 3-60 weeks; (b) animals castrated for 7 days before injection with 5 alpha-dihydrotestosterone (17 beta-hydroxy-5-alpha-androstan-3-one), testosterone and 5 alpha-androstane-3 beta, 17 beta-diol for up to 10 days; (c) rats injected with the 3 beta, 17 beta-diol immediately after castration. Ornithine decarboxylase activity and the concentrations of putrescine, spermidine and spermine were measured. DNA-synthetic activity was monitored by measuring [125I]iododoxyuridine incorporation. An enhanced spermidine/spermine molar ratio reflected increased activity of the prostate. The ratio was higher (greater than 2) in prostates from sexually immature animals, than in the intact adult (1.5), suggesting that the ratio was indicative of the proliferative activity of the tissue. However, in the androgen-stimulated castrated rat, enhanced spermidine/spermine ratios tended to correlate with hypertrophy and secretion. In both sets of experiments there was a linear relationship between protein and spermidine content. High spermidine/spermine molar ratios were the consequence of a relatively low rate of accumulation of spermine relative to spermidine and protein. The relationship between polyamine synthesis and DNA-synthetic activity was investigated in cultured prostate. A combination of insulin (3 mug/ml) and testosterone (0.1 muM caused a stimulatory response in the incorporation of [125I]iododeoxyuridine and in cell division, despite a depleted polyamine content and low ornithine decarboxylase activity in the cultured tissue.

Age Factors↗

The effect of methionine enkephalin on prolactin and luteinizing hormone levels in intact and castrated rats.

The methionine enkephalin (ME) induced responses of prolactin (PRL) and luteinizing hormone (LH) were studied in control and castrated rats bearing chronically-implanted cannulae in the right lateral ventricle. In control rats, 500 microgram ME caused a rise in PRL from a mean +/- SD basal value of 22.8 +/- 12.2 ng/ml to a peak of 195.2 +/- 93.8 ng/ml (p less than 0.001). The peak control occurred 10 min after ME administration and was short-lived. In the same rats, 14 days after castration, there was a significant reduction in both basal (p less than 0.001) and peak PRL response to ME (p less than 0.01), similar to that previously observed with other PRL-inducing stimuli. LH levels, on the other hand, were unaffected by ME treatment in both control and castrated rats.

Animals↗

Notch1-expressing cells are indispensable for prostatic branching morphogenesis during development and re-growth following castration and androgen replacement.

Notch expression is frequently associated with progenitor cells, and its function is crucial for development. Our recent work showing that Notch1 is selectively expressed in basal epithelial cells of the prostate and higher Notch1 expression during development suggests that Notch1-expressing cells may define progenitor cells in the prostate. To test this hypothesis, we have generated a transgenic mouse line in which the Notch1-expressing cells can be ablated in a controlled manner. Specific targeting was achieved by expressing the bacterial nitroreductase, an enzyme that catalyzes its substrate into a cytotoxin capable of inducing apoptosis, under the Notch1 promoter. Cell death in transgenic prostate was confirmed by histological analyses including terminal dUTP nick-end labeling and caspase 3 immunocytochemical staining. We evaluated the consequences of ablation of Notch1-expressing cells in two systems, organ culture of early postnatal prostates and re-growth of prostate in castrated mice triggered by hormone replacement. Our data show that elimination of Notch1-expressing cells inhibited the branching morphogenesis, growth, and differentiation of early postnatal prostate in culture and impaired prostate re-growth triggered by hormone replacement in castrated mice. Furthermore, we found that Notch1 expression following castration and hormone replacement was concomitant with known basal cell markers p63 and cytokeratin 14 and was high in the proliferative human prostate epithelial cells. Taken together, these data suggest that Notch1-expressing cells define the progenitor cells in the prostatic epithelial cell lineage, which are indispensable for prostatic development and re-growth.

Androgens↗

Effects of mepartricin, a polyene macrolide antibiotic, on estrogen-induced hyperplastic growth of the dorsolateral prostate and seminal vesicle in immature castrated rats.

OBJECTIVE: To investigate the effects of mepartricin, a polyene macrolide antibiotic, on estrogen-induced hyperplastic prostate and seminal vesicle (SV) growth in castrated rats. MATERIAL AND METHODS: Immature rats aged 3 weeks were castrated and left untreated for 1 week. Then, 17beta-estradiol benzoate (E(2)-BA) was subcutaneously injected at a dose of 10 microg/day twice weekly, and mepartricin was orally administered at doses of 2.5, 5 and 10 mg/kg once daily for 3 weeks. The weights and hydroxyproline contents of the prostate and SV, the activity of growth factors (GFs) in the dorsolateral prostate (DLP) and the serum estrogen level were measured. Histological examination of the prostate and SV was also performed. RESULTS: Mepartricin dose-dependently suppressed the increase in the serum estrogen level, the weights and hydroxyproline contents of the DLP and SV and the elevation of GF activity in the DLP induced by E(2)-BA treatment. Histological examination also revealed that treatment with mepartricin reduced collagen accumulation and thickening of the smooth muscle layer in the DLP and SV, and proliferation of the glandular epithelium in the DLP. CONCLUSIONS: These results suggest that mepartricin suppresses hyperplastic growth of the DLP and SV induced by estrogen in immature castrated rats, the underlying mechanism being a reduction in the serum estrogen level, thereby suppressing stromal cell proliferation and activation.

Animals↗

Influence of perinatal androgenization on the castration response of adult rats.

An unexplained dichotomy exists between the LH (luteinizing hormone) responses to castration of male and female rats, as males show a more prompt increase in serum LH levels. We have tested the hypothesis that neonatal exposure to androgen determines the sexual dimorphism of that response. Control groups of male and female rats were castrated at 60 days of age. Other animals had been castrated at 0 or 25 days of age and then given steroid treatment via testosterone (T) implants from 25 through 60 days of age. At 60 days of age a blood sample was taken from each animal before removal of either the T implant or the gonads. Animals were bled again 24 and 48 h later. Within 24 h after orchidectomy the typical early plateau of plasma LH had occurred, represented by an increment in mean LH concentrations of 316 ng/ml. Orchidectomy at 25 days of age had little or no effect on subsequent response to removal of T. In contrast, neonatal orchidectomy resulted in a markedly diminished response to T removal on Day 60. The response, however, was not reduced to that of normal females. In female rats plasma LH does not increase by 48 h after ovariectomy. Perinatal testosterone propionate (TP) treatment of females partially masculinized (enhanced) the LH response to T implant removal, but only if ovariectomy had been performed prior to puberty (at 0 or 25 days of age).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Influence of castration of the neonatal rat on the contractile effects of barium chloride and adrenaline in isolated vas deferens: the role of calcium.

Time-response curves to maximal concentrations of barium chloride (BaCl2) (3 X 10(-2) M) and adrenaline (10(-4) M) were studied in vasa deferentia from 3-month-old rats castrated at birth. Either barium or adrenaline was left in the organ baths for 5-min periods, at intervals of about 30 min, and the corresponding isotonic contractions recorded. Two types of effects were measured: the fade response (Jurkiewicz et al., 1977) and the rate at which responses were reduced after Ca2+ withdrawal from nutrient solution. The fade response for BaCl2 was strikingly greater than that in controls. When calcium was removed from the nutrient solution, an almost complete loss of the response to BaCl2 was achieved in less than 3 min for preparations of 3-day castrates, in about 40 min for the organs of 15-day castrates, and in more than 140 min for normal preparations. Treatment with testosterone, 1 week before the experiments, abolished the fade response to BaCl2 and antagonized the loss of responsiveness observed for this substance in a calcium-deficient solution. These data suggest that the production of testosterone by the testis during the critical period of neonatal differentiation is important for the translocation of calcium ions in the isolated vas deferens of the adult rat.

Animals↗