Changes in the hypothalamic contents of LHRH-I and -II and in pituitary responsiveness to synthetic chicken LHRH-I and -II during the progesterone-induced surge of LH in the laying hen

The contents of LHRH-I and -II in the anterior hypothalamus and posterior hypothalamus (including the mediobasal hypothalamus and median eminence) were measured at 90, 180 and 360 min after the i.m. injection of laying hens with progesterone. Whilst no changes were observed in the content of LHRH-I in the anterior hypothalamus, LHRH-I in the posterior hypothalamus tended to fall at 90 and 180 min after injection of progesterone in hens maintained on 16 h light:8 h darkness (16L:8D) and 8L:16D respectively. Pretreatment of laying hens with tamoxifen significantly increased the hypothalamic contents of LHRH-I and -II, raised the basal plasma concentration of LH and modified the LH response to progesterone injection. In hens in which tamoxifen prevented an increase in the plasma concentration of LH after progesterone injection, the content of LHRH-I in the posterior hypothalamus remained unchanged. In contrast, in hens in which progesterone stimulated a steep increase in LH within 90 min, there was a pronounced and significant fall in LHRH-I content of the posterior hypothalamus. No change in the hypothalamic content of LHRH-II was observed during the progesterone-induced surge of LH until plasma concentrations had attained maximal values or started to decline. Then, in hens maintained on 16L:8D, a significant fall in the content of LHRH-II in the anterior hypothalamus was found at both 180 and 360 min after injection with progesterone. Tests in vitro and in vivo of the responsiveness of the pituitary gland to synthetic LHRH-I and -II revealed no change at 90 min after injection of laying hens with progesterone, when plasma concentrations of LH were increasing, but a pronounced reduction when plasma LH concentrations were maximal or falling. These results suggest that LHRH-I mediates in the progesterone-induced increase in the plasma concentration of LH. Although the subsequent decline in plasma LH was associated with a reduced responsiveness of the pituitary gland to LHRH, a significant correlation between the contents of LHRH-I and -II in the anterior hypothalamus and a fall in the hypothalamic content of LHRH-II when plasma LH was maximal or declining allows the possibility of an involvement of this peptide in the neuroendocrine events preceding ovulation.     

Absence of an effect of naloxone, an opioid antagonist, on luteinizing hormone release in vivo and luteinizing hormone-releasing hormone I release in vitro in intact, castrated, and food restricted cockerels

The possibility that the tonic secretion of luteinizing hormone (LH) and chicken luteinizing hormone-releasing hormone I (LHRH-I) is regulated by an inhibitory action of endogenous opioid peptides was investigated in cockerels using the opiate receptor antagonist, naloxone. Baseline concentrations of plasma LH in the experimental cockerels were increased by surgical castration or reduced by limiting food intake. Baseline and K(+)-induced releases of LHRH-I from perifused mediobasal-preoptic hypothalami from castrated cockerels were higher than those from hypothalami from intact cockerels. Similarly, baseline and K(+)-induced releases of LHRH-I from perifused mediobasal hypothalami from fully fed cockerels were higher than those from the hypothalami from fasting cockerels. Intravenous injections of 0.1, 1, or 10 mg naloxone/kg body weight failed to increase the concentration of plasma LH in castrated, intact, fully fed, or fasted cockerels. Perifusion of mediobasal-preoptic hypothalami from castrated or intact cockerels with 200 microM naloxone or mediobasal hypothalami from fully fed or fasted cockerels with 10 microM naloxone failed to stimulate the release of LHRH-I. These observations suggest in the cockerel that endogenous opioid peptides may not play an obligatory role in the inhibitory control of the tonic secretion of luteinizing hormone.     

Diurnal changes in the plasma concentrations of LH and hypothalamic contents of LHRH-I and LHRH-II in the domestic hen

Diurnal changes of LH secretion in sexually immature hens of 9, 11, 13 and 15 weeks of age consisted of 25-40% increases in the mean concentrations of LH in plasma between 15.00 and 18.00 h, i.e. between 2 h before and 1 h after the onset of darkness. During this time there was a tendency for the mean contents of LHRH-I in the anterior hypothalamus and posterior hypothalamus to increase by 21-74% and 20-56% respectively. In hens of 9 and 15 weeks, diurnal changes in the plasma concentration of LH closely paralleled those of LHRH-I content in the posterior hypothalamus. In contrast, the diurnal rhythm of LH secretion in hens of 11 and 13 weeks was more marked and plasma concentrations of LH continued to rise steeply between 18.00 and 21.00 h, i.e. between 1 and 4 h after the onset of darkness. At 11 weeks, this was associated with a reduction (P less than 0.01) in the contents of LHRH-I and LHRH-II, particularly in the anterior hypothalamus. In laying hens, a diurnal decline (P less than 0.01) in the plasma concentration of LH between 1 and 4 h after the onset of darkness was preceded by a fall (P less than 0.05) in the content of LHRH-I in the posterior hypothalamus and in the total hypothalamic content of LHRH-II (P less than 0.01).(ABSTRACT TRUNCATED AT 250 WORDS)     

Pituitary receptor sites for gonadotropin-releasing hormone: effect of castration and substitutive therapy with sex steroids in the male rat

To examine the role of pituitary gonadotropin-releasing hormone (GnRH) receptors (pit GnRH-R) in the regulation of gonadotropin secretion, male rats were orchidectomized and then selectively received substitutive therapy with sex steroids. Pituitary content of GnRH-R was determined by saturation analysis, using radioiodinated [D-Trp6,(N-Et)Pro5,des-Gly10]GnRH as tracer. Castration produced a rapid and sustained increase of the number of GnRH-R, which doubled after 2 days, and after 10 days the pituitary content of GnRH-R was 258 +/- 23 fmol/pituitary compared to 103 +/- 12 fmol/pituitary for sham-operated control animals. No change of the affinity constant (Ka) was observed (Ka = 1.13 +/- 0.08 X 10(10) M-1; n = 14). Plasma LH increased 5- to 10-fold and FSH-2- to 3-fold after castration, and hypothalamic GnRH content was depleted by 30-60%. Immediate substitution of castrated rats with testosterone propionate (250 micrograms daily) prevented the increases of both plasma gonadotropins and of GnRH-R. Treatment of acutely castrated rats for 7 days with testosterone propionate (50-200 micrograms), 5 alpha-dihydrotestosterone propionate (25-400 micrograms), or estradiol benzoate (2 micrograms) prevented the rise in pit GnRH-R in a dose-related manner and normalized the other parameters studied except that plasma FSH remained slightly elevated. In contrast, when substitutive therapy was started 8 days after castration or later, the 7-day treatment with sex steroids reduced plasma gonadotropins, but pit GnRH-R remained elevated, and hypothalamic GnRH content remained depleted. These results indicate that the marked increase of gonadotropin secretion after castration is mediated at least in part, by an increase in the number of pit GnRH-R. Sex steroids were able to reverse all castration-induced endocrine changes in acutely castrated rats, but in long term castrated animals their action at higher centers to normalize hypothalamic GnRH content, and indirectly, to reduce pit GnRH-R content, was either delayed or ineffective. Thus, the rapid feedback action of sex steroids in long term castrated rats may be predominantly exerted at the pituitary level.     

Increased sensitivity to the negative feedback effects of testosterone induced by hyperprolactinemia in the adult male rat

High plasma levels of PRL induced by transplants of two donor pituitaries under the kidney capsule of adult male rats resulted in a prolonged suppression of plasma levels of LH and FSH although testosterone levels were maintained within normal limits. Castration of rats with pituitary transplants resulted in a normal though delayed rise in serum levels of both LH and FSH to levels equivalent to those in normal castrated controls. This increase in gonadotropin levels occurred in spite of maintenance of elevated PRL levels. Two experiments were carried out in which testosterone was restored after castration by Silastic testosterone-containing implants of various lengths (Exp 1:60, 30, and 10 mm; Exp 2: 30, 20, 10, 5, and 2 mm). In both experiments 60- and 30-mm testosterone implants prevented the postcastration rise in LH and FSH in both control and hyperprolactinemic rats. However, although the shorter testosterone implants delayed this rise in control rats, levels of LH and FSH increased by 4 days and were not significantly different from castrated rats without testosterone implants by 15 days after castration. In contrast, this rise in gonadotropins was abolished or considerably delayed by the shorter implants in hyperprolactinemic rats, demonstrating an increase in sensitivity of the hypothalamic pituitary axis to the negative feedback effects of testosterone in these animals. These results suggest that 1) to maintain suppression of gonadotropin secretion in hyperprolactinemia high levels of PRL alone are insufficient and gonadal steroids are required, and 2) high levels of PRL appear to sensitize the hypothalamic-pituitary axis to the negative feedback effects of gonadal steroids.     

Androgenic and oestrogenic steroid participation in feedback control of luteinizing hormone secretion in male sheep

The acute castrate ram (wether) was used as an experimental model to investigate the site(s) of feedback on luteinizing hormone (LH) by testosterone, dihydrotestosterone and oestradiol. At the time of castration, wethers were implanted subdermally with Silastic capsules containing either crystalline testosterone (three 30 cm capsules), dihydrotestosterone (five 30 cm capsules) or oestradiol (one 6.5 cm capsule). Blood samples were taken at 10 min intervals for 6 h 2 weeks after implantation to determine serum steroid concentrations and to characterize the patterns of LH secretion. Pituitary LH response to exogenous LRH (5 ng/kg body weight) were also determined at the same time. The steroid implants produced serum concentrations of the respective hormones which were either one-third (testosterone) or two-to-four times (dihydrotestosterone, oestradiol) the levels measured in rams at the time of castration. Non-implanted wethers showed rhythmic pulses of LH (pulse interval 40-60 min) and had elevated LH levels (16.1 +/- 1.6 ng/ml; mean +/- SE) 2 weeks after castration. All three steroids suppressed pulsatile LH release and reduced mean LH levels (to below 3 ng/ml) and pituitary LH responses to LRH. Inhibition of pulsatile LH secretion by all three steroids indicated that testosterone as well as its androgenic and oestrogenic metabolites can inhibit the LRH pulse generator in the hypothalamus. Additional feedback on the pituitary was indicated by the dampened LH responses to exogenous LRH.     

The in-vitro transformation of [3H]dehydroepiandrosterone into its principal metabolites in the adrenal cortex of adult castrated male rats and following steroid treatment

The adrenal gland of castrated adult male rats metabolized [3H]dehydroepiandrosterone in vitro to delta 4-androsten-3,17-dione (4AD), testosterone, dihydrotestosterone (DHT) and 5 alpha-androstane-3,17-dione (5 alpha AD). Despite the low testosterone values, DHT and 5 alpha AD were higher 30 and especially 60 days after castration, with raised 4AD:testosterone and decreased testosterone:DHT ratios. The 5 alpha-reductase activity thus appears to increase with time after castration. Fourteen days after castration, 4AD was the only metabolite that was raised compared with intact animals, and testosterone was comparable in sham-operated and castrated rats. The administration of testosterone propionate to castrated rats restored testosterone values to those of intact rat adrenals, whereas 4AD values were greater. The administration of dihydrotestosterone propionate also yielded higher levels of 4AD, in the presence of a lower testosterone value. After administration of oestradiol benzoate, 4AD values were lower especially compared with the other hormone-treated groups, and there was an unexpectedly high testosterone value. These data indicate that the adrenal gland contributes to the production of androgens, as previously noted by Andò, Canonaco, Beraldi et al. (1988) who showed increased plasma 4AD and testosterone levels in adult male rats 30 days after castration. Furthermore, adrenal androgen production in castrated animals is differentially regulated by sex steroids.     

The effect of ovarian steroids on hypothalamic norepinephrine neuronal activity

The concentration of norepinephrine and the apparent turnover of norepinephrine in the hypothalamus was studied in castrated rats and after the injection of ovarian steroids. Both parameters were found to vary not only with the hormone treatment, but also with the time of the day. Norepinephrine concentration in the anterior hypothalamus of castrated male and female rats was significantly higher in the afternoon than in the morning, but no statistical difference was observed in the concentration of norepinephrine in the posterior hypothalamus. Three days after the injection of a single dose of 20 microgram oestradiol benzoate, norepinephrine concentration and apparent turnover were found with decreased significant values in the anterior hypothalamus of castrated females, killed in the afternoon. Progesterone 2 mg injected three days after the priming dose of oestradiol benzoate, produced 4 h later in the afternoon a significant rise in norepinephrine concentration and apparent turnover. These results suggest that the apparent turnover of hypothalamic norepinephrine in the afternoon is reduced by oestrogen and increased by oestrogen plus progesterone.     

Differential effects of testosterone, 5 alpha-dihydrotestosterone and oestradiol-17 beta on plasma concentrations of LH in castrated ferrets

The biological activity of testosterone often depends on the conversion of testosterone within the target cell to an androgenic or oestrogenic metabolite. The purpose of this study was to compare the relative ability of testosterone and two of its metabolites, dihydrotestosterone (DHT) and oestradiol, to suppress LH secretion in castrated male ferrets. Castrated ferrets were treated with five different doses of steroid by implanting various numbers of s.c. silicone elastomer capsules packed with either testosterone, DHT or oestradiol. The lowest dose of oestradiol (0.1 mm capsule length/100 g body weight, mean estimated total release rate of 25 ng/day) significantly suppressed plasma concentrations of LH in castrated ferrets. Higher amounts of DHT (2.5 mm capsule length/100 g body weight, mean estimated total release rate of 88 ng/day) were required for a significant reduction in plasma concentrations of LH. Concentrations of LH were also significantly lowered by testosterone when administered at a 2.5 mm capsule length/100 g body weight; however, estimated total release rate was 312 ng/day from these capsules. The fact that oestradiol was more effective than DHT, and that DHT was more effective than testosterone in inhibiting LH secretion in castrated ferrets, suggests that in gonadally intact ferrets, testosterone may be converted to DHT or oestradiol within target cells that mediate steroid negative feedback on LH secretion.     

Concentrations of immunoreactive luteinizing hormone releasing hormone in discrete brain regions of the cockerel: effects of castration and testosterone replacement therapy

The validity of using a radioimmunoassay employing an antiserum raised against synthetic luteinizing hormone releasing hormone (LH-RH) for the quantification of luteinizing hormone releasing factor (LH-RF) in birds was investigated. Extracts of avian hypothalamus yielded displacement curves which were parallel to that of the synthetic LH-RH standard and the immunoreactive potencies of a number of extracts assayed concurrently using two different anti-LH-RH sera were found to be similar. Moreover, after chromatography of cockerel hypothalamic extract on carboxymethyl-cellulose, immunoreactive and biologically active LH-RF were found in the same eluate fractions. Immunoreactive LH-RH was shown to be widely distributed in cockerel hypothalamus with the highest concentrations present in the mediobasal hypothalamus (MBH; 6.55 +/- 1.86 pg/microgram protein, n = 6) and medial preoptic region (POR; 0.95 +/- 0.07 pg/microgram protein, n = 6). The postcastration rise in plasma LH in the cockerel was accompanied by significant (P less than 0.05) increases in the concentration of LH-RH in five hypothalamic areas including the POR; testosterone replacement therapy completely reversed these effects. Although castration raised the mean concentration of LH-RH in four other hypothalamic areas including the MBH, these differences were not significant. However, testosterone replacement therapy depressed LH-RH in all four regions to levels significantly (P less than 0.05) less than those in castrated cockerels. These findings constitute the first direct evidence that the negative feedback action of testosterone on LH secretion in the cockerel is mediated, at least in part, by an action on hypothalamic LH-RF-producing neurones.     

Testosterone raises neuropeptide-Y concentration in selected hypothalamic sites and in vitro release from the medial basal hypothalamus of castrated male rats

Although neuropeptide-Y (NPY)-containing neurons are widely distributed in the hypothalamus, castration decreased NPY concentrations only in the median eminence (ME), arcuate nucleus (ARC), and ventromedial nucleus (VMN). We have now examined the effects of testosterone (T) replacement in 2-week castrated male rats on NPY levels in hypothalamic and preoptic area regions and in vitro NPY release in three experiments. In the first experiment we studied the effect of T on NPY concentration in castrated rats. Two-week castrated rats were implanted sc with T-filled or empty Silastic capsules 30 mm in length. Ten days later rats were killed, and NPY levels were measured by RIA in microdissected sites. T implants raised serum T levels to the range found in gonad-intact rats and decreased serum LH levels to the basal range. Further, of the six brain sites examined, significant increases in NPY concentrations occurred selectively in the ME, ARC, and VMN of T-implanted rats. In the second experiment, the ability of T to reverse the effect of castration on NPY levels compared to those in intact (sham) rats was assessed. Again, castration decreased NPY levels in the ME, ARC, and VMN only, and replacement of physiological levels of T restored NPY levels approximately 100%, 127%, and 74% in the ARC, VMN, and ME, respectively. In the third experiment, the effect of castration and T implants (30-mm T capsules for 10 days) to 2-week castrated rats on the in vitro release of NPY from medial basal hypothalamus (MBH) was assessed. Basal NPY release was not significantly changed after castration and T replacement. However, in response to a 30-min pulse of KCl (45 mM) NPY release from the MBH of castrated rats was significantly reduced compared to that in intact and T-replaced castrated rats. These studies show that castration decreases and T replacement restores NPY levels selectively in three hypothalamic sites, viz. ME, ARC, and VMN, and KCl-induced NPY release from the MBH in vitro is decreased after castration and restored by T replacement, thereby suggesting that a local subset of androgen-concentrating neurons may regulate NPY levels and release in a site-specific manner. Further, these results are in line with our emerging view that gonadal steroids modulate neurosecretion not only of LHRH, but also of other functionally linked regulatory peptides.     

Effects of anterior hypothalamic implantation of gonadal steroids on serum and pituitary gonadotrophins (follicle-stimulating hormone and luteinizing hormone) in the castrated male rat.

Castrated rats were stereotaxically implanted with 1 microng oestradiol benzoate, 5 microng testosterone isobutyrate or, as a control, 10 microng cholesterol, in the hypothalamus. The effects of the steroids on plasma and pituitary gonadotrophins (FSH and LH) were assessed by radioimmunoassay. Our results indicate that, in the male rat, in addition to the arcuate nucleus-median eminence complex, the preoptic suprachiasmatic area is able to control synthesis and secretion of both gonadotrophins, and that it is sensitive to oestradiol and testosterone.     

Effects of oestradiol on gonadotrophin levels in normal and castrated men

Context  Testosterone inhibits gonadotrophin release in men either directly or after aromatization to oestradiol. We hypothesized that in males the androgen receptor-mediated effect of testosterone on LH release is negligible relative to that of oestradiol.
Objective  To compare the effect of experimentally induced variations of plasma oestradiol levels on LH levels in normal (physiological testosterone levels) and castrated men (very low testosterone levels).
Design  Prospective, open label, intervention.
Subjects and interventions  We suppressed endogenous oestradiol in 10 young men with letrozole 2·5 mg once daily. In these men and in 10 young healthy castrated men, we restored plasma oestradiol levels with oestradiol patches (first week 100 μg/day, second week 50 μg/day, third week 25 μg/day and fourth week no oestradiol patch).
Measurements  The effect of the intervention on plasma levels of LH were monitored and compared between the groups.
Results  With the intervention, the mean plasma oestradiol level in the two groups varied from supraphysiological to below the lower reference range. Levels of LH mirrored plasma oestradiol levels in both the groups, as did testosterone in the intact group. Despite similar oestradiol levels, mean levels of LH were significantly higher in the castrated group compared to the intact group for all doses of oestradiol, and supraphysiological levels of oestradiol were unable to suppress LH into the physiological range in the castrated group.
Conclusions  Physiological plasma oestradiol levels have a substantial suppressive effect on LH in men. However, low-normal testosterone levels are a prerequisite for suppression of LH into the normal range.     

Steroidal regulation of hypothalamic neuropeptide Y release and gene expression.

Neuropeptide Y (NPY) readily stimulates the release of hypothalamic LHRH and pituitary LH release in intact and gonadal steroid-primed gonadectomized rats. We have now tested the hypothesis that the release and synthesis of hypothalamic NPY may be regulated by gonadal steroids. To measure the effects of gonadal hormones on NPY release, a permanent push-pull cannula was implanted in the anterior pituitary (AP) of sham castrated (controls) or castrated (CAST) male rats, and 1 week later, the AP was perfused with artificial cerebrospinal fluid over a 3-4 h period. NPY concentrations in the perfusates collected at 10-min intervals were measured by RIAs. The NPY release pattern in the AP was episodic in both intact and CAST rats, and the frequency of NPY episodes was similar in two groups. However, the amount of NPY detected in the AP of CAST rats was significantly less than that of intact rats because the mean rate of release and the amplitude of NPY episodes in the perfusates of CAST rats were significantly reduced. This observation of attenuated hypothalamic NPY output in vivo and previous evidence of decreased hypothalamic NPY contents after CAST implied that the synthesis of hypothalamic NPY may be regulated by testicular secretions. Therefore, the effects of testosterone (T)-replacement on preproNPY messenger RNA (mRNA) in the medial basal hypothalamus (MBH) was evaluated. Rats were CAST and received either empty or T-filled Silastic capsules sc. Two weeks later, the level of perproNPY mRNA in the MBH was determined by solution hybridization/ribonuclease protection assay using a complementary RNA probe complementary to the rat NPY precursor mRNA. We observed that the levels of preproNPY mRNA were 2-fold higher in the MBH of T-replaced CAST as compared to control CAST rats. These findings are consistent with the hypothesis that gonadal steroids enhance the neurosecretory activity of hypothalamic NPYergic neurons, and for the first time reveal a coupling between the level of gene expression and the secretion of a neuropeptide involved in the regulation of hypothalamic LHRH and pituitary LH release.     

Effects of castration and gonadal steroids on serum luteinizing hormone and prolactin in old and young rats.

Changes in serum LH and prolactin concentrations in response to bilateral gonadectomy and gonadal steroid replacement were measured in mature young (4-6 months) and old (23-30 months) female and male Long-Evans rats. On day 13 after gonadectomy, female rats were injected with oestradiol benzoate (OB) and male rats with testosterone propionate (TP) for a period of 12 days. They were then permitted a recovery period of 6 weeks. Serum prolactin and LH concentrations were measured by radioimmunassay in single blood samples taken at various intervals before and after gonadectomy and during and after steroid treatment. Serum LH levels were about the same in intact young old female rats, but after ovariectomy LH rose several fold higher in young than in old remale rats. In male rats, after orchidetomy the increase in serum LH was greater in young than in old rats. Oestradiol benzoate and TP injections into female and male young and old rats produced variable effects on LH release. Serum prolactin concentrations were approximately six times higher in old intact than in young intact female rats, and after ovariectomy showed a much greater percentage reduction in old than in young female rats. Administration OB produced a greater absolute increase in serum prolactin in old than in young female rats. Serum prolactin values were about the same in old and young male rats, and the effects of castration and TP administration on serum prolactin were not markedly different in the two age groups. These results indicate that old female and male rats are less capable of releasing LH than young rats of both sexes, but old females release more prolactin than young females.     

Effects of anti-oestrogen treatment of neonatal male rats on lordosis behaviour and mounting behaviour in the adult

Male rats were treated daily with 100 microgram of the anti-oestrogen ethamoxytriphetol (MER-25) or oil during the first 10 days of life and tested for lordosis behaviour and mounting behaviour as intact adults, after castration and after castration and oestradiol benzoate or testosterone propionate treatment. The MER-25-treated rats showed higher levels of lordosis behaviour than oil-treated rats in all four treatment groups. Under each of these endocrine conditions, except after castration alone, the MER-25-treated rats showed a reduced capacity to ejaculate. Treatment of the neonatal rat with MER-25 reduced body weight in adulthood but did not change the weight of the accessory sexual glands, the testes, the number of cornified papillae on the glans penis or plasma testosterone concentrations during development. The response of the accessory sexual glands and cornified papillae on the glans penis to treatment with oestradiol benzoate or testosterone propionate after castration in adulthood was unaffected by treatment with MER-25. It is suggested that formation of oestrogen in the neonatal male rat brain from testosterone in the circulation inhibits the capacity to show lordosis behaviour and facilitates the capacity to ejaculate in response to ejaculate in response to gonadal hormone treatment in adulthood.     

Effect of testosterone on gonadotrophin-releasing hormone receptors in the castrated rat: preliminary evidence for a stimulatory effect of testosterone on gonadotrophin function in the male rat

In the long-term castrated rat the negative feedback effect of testosterone is markedly reduced and the raised levels of plasma LH seen in the castrated animals are not suppressed by physiological concentrations of plasma testosterone. In this study we have measured pituitary gonadotrophin-releasing hormone (GnRH) receptor content as well as plasma and pituitary LH on days 1, 10 and 40 after castration and noted the effect of testosterone replacement on these parameters. We found that the negative feedback effect of physiological concentrations of testosterone on plasma and pituitary LH, pituitary GnRH receptor content and response to exogenous GnRH was attenuated 10 and 40 days after castration. It is suggested that the lack of effect of testosterone in the long-term castrated rat is due to its inability to reduce the pituitary GnRH receptor content. On increasing testosterone to supraphysiological levels, the negative feedback effect was reinstated. We also found that in rats 40 days after castration, physiological and subphysiological concentrations of testosterone significantly increased pituitary GnRH receptor content and this may explain the previous findings that low concentrations of testosterone can enhance the effect of GnRH and increase plasma LH levels.     

Further studies on the effects of testosterone on hypothalamic LH-RH and serum LH levels: castration-induced delayed response

We have previously reported that luteinizing hormone releasing hormone (LH-RH) levels in the medial basal hypothalamus (MBH) of adult male rats castrated for 2 weeks can be raised by testosterone (T) or estradiol (E2) treatment for 3-4 days. The present study was undertaken to determine the time after castration when the hypothalamus becomes refractory to this feedback action and whether prolongation of exposure to steroids would reinstate the hypothalamic LH-RH response. Treatment with T (Silastic capsules, s.c.) for 4 days, when commenced either immediately or 14 days after castration, significantly raised the MBH LH-RH levels. A 4-day regimen of E2 or dihydrotestosterone, on the other hand, evoked similar increase when commenced up to 21-28 days after castration. Further delay in institution of the 4-day treatment rendered the three steroids completely ineffective. When the effects of prolongation of treatment were tested in long-term castrated rats (56 days), it was found that T required 14 days as compared to 7 days of exposure with E2 or dihydrotestosterone to produce an equivalent MBH LH-RH response. Furthermore, the MBH LH-RH and serum LH responses were not always correlated, since increments in the MBH LH-RH were observed when serum LH levels were suppressed by physiological levels of T or dihydrotestosterone and also when they were unchanged with low levels of T or E2. These studies show that long-term castration causes a dichotomy in responsiveness of the hypothalamo-pituitary axis to steroids.(ABSTRACT TRUNCATED AT 250 WORDS)     

Plasma vasopressin and oxytocin levels in intact goats and in castrated goats given testosterone

Oxytocin, vasopressin, cortisol and testosterone levels in the plasma were measured by radioimmunoassay in intact male goats as well as in prepubertally castrated goats injected daily, for 2 weeks, with oil vehicle and then, for 4 weeks, with testosterone propionate in oil to study the influence of gonadal steroids on posterior pituitary hormones. Packed cell volume, plasma osmolality and sodium concentration were also measured in all blood samples. Plasma levels of oxytocin, vasopressin and cortisol were similar in the intact and oil-injected castrated goats. Testosterone treatment significantly increased plasma levels of oxytocin (P less than 0.01) in castrated goats but the increased levels were similar to those seen in the intact goats at the same time of year. Plasma levels of cortisol and vasopressin were unaffected by testosterone propionate treatment, whereas packed cell volume was significantly decreased (P less than 0.01). Testosterone treatment of castrated male goats appears not to have any action on pituitary hormones and oxytocin increases in the spring in both intact and castrated male goats.     

Effects of functional and surgical castration of white leghorn cockerels, and replacement therapy, on food intake, obesity, reproductive traits, and certain components of blood, liver, muscle, and bone

Basal hypothalamic bilateral electrolytic lesions were placed in White Leghorn cockerels causing functional castration and obesity. Other cockerels were surgically castrated. Determinations were made of the reproductive traits, weight of abdominal adipose tissues, weight and fat content of livers, fat in muscles and bones, hematocrits, plasma calcium, triglycerides, cholesterol, and phospholipid phosphorus before and after injections of chicken pituitaries (CP) into cocks bearing hypothalamic lesions (HL), and before and after injections of testosterone propionate (TP) into surgically castrated (SC) cocks. Treatment of HL cocks with CP partially restored reproductive behavior, but abdominal massage produced only seminal fluid without spermatozoa. The weight of the abdominal adipose tissue was used as a measure of adiposity since it varies directly with total body fat. Adiposity in HL cocks was 2.5 times greater than that of SC cocks. Injections of CP into HL cocks reduced adiposity by 60%. In contrast, injections of TP into SC cocks reduced body fat to normal. Injections with CP into HL cocks reduced moderately the weights of the hypertrophied fatty livers and reduced fat content toward normal, but livers remained abnormal. In surgically castrated cocks, livers remained normal. Functional or surgical castration depressed blood hematocrit values. Injections with TP restored hematocrit values in SC cocks. In contrast, injections of CP into HL cocks were virtually without effect upon the depressed hematocrit values.