Age-related discrepancies between serum and pituitary gonadotrophin, and pituitary gonadotrophin subunit mRNA responses to castration and testosterone replacement in male rats

1992 ◽  
Vol 135 (3) ◽  
pp. 507-515 ◽  
Author(s):  
P. Pakarinen ◽  
I. Huhtaniemi

ABSTRACT The responses of gonadotrophin gene expression, pituitary content and serum levels to castration alone and castration plus testosterone replacement (silicone elastomer implants) were compared in male rats at 10, 30, 60 and 90 days of age. Sham-operated animals served as controls. In addition, 30-day-old castrated rats were treated with dihydrotestosterone (DHT) and diethylstilboestrol (DES). When killed 7 days after castration, the increases in serum LH (six- to eightfold; P < 0·01) and FSH (two- to fourfold; P < 0·01) were similar at all ages studied. Likewise, testosterone reversed the effects of castration in a largely similar fashion at all ages. In contrast, great age-related differences were observed in the responses of gonadotrophin subunit mRNAs to the treatments. Castration increased the common α subunit mRNA two- to fourfold on days 10 and 30 (P < 0·01), sixfold on day 60 (P < 0·01), but not at all on day 90. Testosterone reversed the increases at all ages, but the levels were below those of controls only at 90 days (P < 0·01). The highest increases (sixfold; P < 0·01) of LH-β mRNA were seen on days 10 and 60, the others being two- to threefold higher (P < 0·05–0·01). Testosterone reversed this effect at 60 days and suppressed LH-β mRNA to below the control levels at other ages (P < 0·01). Castration had no effect on FSH-β subunit mRNA at 30 and 90 days but a four-to fivefold increase was seen on days 10 and 60 (P < 0·01). Testosterone suppressed these mRNAs at all ages, and they decreased to below the levels in controls at 30 and 60 days. Testosterone, DHT and DES had, at 30 days, practically the same effects on the LH parameters, whereas DHT was clearly less effective than testosterone and DES in suppressing those of FSH. In conclusion, although there was, in general, good agreement between gonadotrophin mRNA and serum levels in response to castration and testosterone replacement, there were specific ages when the post-castration increases in FSH and/or LH occurred with no detectable change in the respective mRNA levels. These findings indicate that altered transcription (or mRNA stability) is not solely responsible for the responses of the gonadotrophins to altered gonadal feedback, but that changes in translation efficiency and/or serum gonadotrophin stability are involved at specific ages of development. Journal of Endocrinology (1992) 135, 507–515

1988 ◽  
Vol 1 (1) ◽  
pp. 61-68 ◽  
Author(s):  
S. D. Abbot ◽  
K. Docherty ◽  
R. N. Clayton

ABSTRACT The control of anterior pituitary hormone gene expression by testosterone in male rat pituitaries in vivo was investigated using dot-blot mRNA-cDNA hybridization assays. Common α subunit mRNA levels doubled by 2 days after orchidectomy and rose progressively to reach plateau levels three to four times intact control values by 2 weeks. LH-β mRNA increased significantly (≃50%) within 12h, and thereafter progressively to seven times intact control values by 3 weeks after orchidectomy. The changes in α mRNA were likely to have occurred in gonadotrophs and not thyrotrophs, since TSH-β mRNA levels were unaltered by orchidectomy. LH subunit mRNA changes were accompanied by an initial (1–4 days) decrease in pituitary LH content; thereafter, pituitary LH increased in parallel with and by a similar magnitude to the LH-β mRNA. Serum LH rises occurred before significant increases in LH subunit mRNA after orchidectomy. The lack of temporal correlation between mRNA levels and serum and pituitary LH in the early stages after removal of testosterone feedback contrasts with the good correlation when a new steady state was achieved after 3–4 weeks, and indicates differing kinetics for changes in these aspects of gonadotroph function. An inhibitory effect of testosterone on LH subunit gene expression was confirmed by prevention of the rise in α and LH-β mRNAs when treatment commenced immediately after castration. However, pituitary LH content and serum LH levels were reduced relative to control values, suggesting additional inhibitory actions of testosterone on translational and post-translational events in gonadotrophs. A stimulatory effect of testosterone on α mRNA levels was observed between 4 and 24 h after a single injection in rats castrated 2 weeks previously, no effect being seen on LH-β mRNA. The mechanism for this action remains to be elucidated. Gene specificity of testosterone action was confirmed by unaltered levels of mRNA for prolactin, GH, TSH-β subunit and actin under all experimental conditions. No changes in pituitary content of prolactin or GH were found. We conclude that regulation of LH subunit gene expression by testosterone is an important step in control of gonadotrophin synthesis and availability for release.


1992 ◽  
Vol 8 (3) ◽  
pp. 249-258 ◽  
Author(s):  
I. S. Scott ◽  
M. K. Bennett ◽  
A. E. Porter-Goff ◽  
C. J. Harrison ◽  
B. S. Cox ◽  
...  

ABSTRACT Hypogonadal (hpg) mutant mice, with a congenital deficiency of hypothalamic gonadotrophin-releasing hormone (GnRH), and testicular feminized (tfm) mice, which lack a functional androgen receptor, were used to study the effects of the potent GnRH agonist 'Zoladex' (ICI 118630; d-Ser (But)6, Azgly10-GnRH) on pituitary and gonadal function. Zoladex (0.5 mg) in a sustained-release lactide—glycolide copolymer depot was administered subcutaneously under anaesthesia and was left in place for 7 days, after which time the effects of the drug upon pituitary and serum gonadotrophin concentrations, glycoprotein hormone subunit mRNAs and testicular morphology were investigated. At the pituitary level, Zoladex treatment resulted in a substantial reduction in LH content in normal males, and LH content was depressed in hpg mice even below the basal levels normally found in these mutants. Pituitary LH content in the Zoladex-treated animals was depressed in the tfm groups, but not to the same levels as those found in the normal and castrated normal mice. Zoladex treatment at the time of castration prevented the post-operative elevation in serum LH associated with castration alone. In the androgen-deficient tfm mouse, Zoladex did not depress the normally elevated serum LH levels. Serum LH in the hpg animals was, in all cases, below the limit of detection of the assay. Pituitary FSH content was depressed into the hpg range in both the normal and castrated animals, but there was no further depression in the hpg mice. The pituitary content was reduced in the tfm mice, again the effects not being as dramatic as in the normal and castrated animals. Serum FSH content, as measured by radioimmunoassay, was depressed by 50% in normal mice; there was no reduction in the hpg mice, however. With regard to pituitary gonadotrophic hormone gene expression, Zoladex administration to normal mice caused a dramatic reduction in LHβ mRNA content, to a level approximating that found in untreated hpg mice. The drug also depressed LHβ mRNA in the castrated group to the hpg range when given at the time of castration, whereas in untreated castrated mice there was a significant increase in LHβ mRNA. In the tfm mouse, which can be considered as a model for long-term failure of androgen feedback, Zoladex again induced a fall in LHβ mRNA, but not to the same extent as in the normal and normal castrated group. Zoladex had no effect on the already low levels of LHβ mRNA found in hpg mice. Pituitary FSHβ mRNA levels were not significantly altered by Zoladex in any of the treatment groups, whereas the drug induced a substantial rise in the common α-subunit mRNA in normal and hpg mice, to a level equalling that found in castrated tfm mice. In the latter two groups, Zoladex treatment did not result in a further increase in α-subunit mRNA above that found after castration alone, or in the untreated tfm mutant. Treatment for 7 days with Zoladex resulted in a significant increase in testis weight, with spermatogenesis advancing beyond the first meiotic division with many round spermatids found within the seminiferous tubules. However, the interstitial cells remained atrophic and there was evidence of seminal vesicle growth. Nevertheless, there was a small but significant increase in testicular androgen content. Administration of the agonist to hypophysectomized hpg mice did not stimulate testicular or seminal vesicle growth, suggesting that the drug does not stimulate steroidogenesis via a direct action upon the testis. Overall, the pharmacological effects of the drug appear to have turned off the transcription of the LHβ gene, with a consequent reduction in LH synthesis and probably also secretion in the longer term. With FSHβ, gene transcription was apparently unchanged and, with a substantial increase in the common α-subunit message, it would appear that the pituitary gland of Zoladex-treated animals may be predominantly biased towards FSH secretion. Although the circulating FSH levels as measured by radioimmunoassay were unaltered by Zoladex, there are several reports that GnRH agonists increase serum levels of bioactive hormones, perhaps by altering glycosylation of the FSH dimer glycoprotein.


1989 ◽  
Vol 121 (3) ◽  
pp. 409-417 ◽  
Author(s):  
M. Bergendahl ◽  
A. Perheentupa ◽  
I. Huhtaniemi

ABSTRACT The effects of 4–6 days of food deprivation on the pituitary-testicular function of adult male rats were studied. Fasting decreased body weights on average by 23% (P<0·01) and those of seminal vesicles by 55% (P<0·01) in 4 days. No consistent changes were found in testicular and ventral prostate weights. The pituitary levels of gonadotrophin-releasing hormone (GnRH) receptors decreased by 50% (P<0·01). Serum and pituitary levels of LH, FSH and prolactin decreased by 25–50% (P<0·01 for all). Testicular and serum levels of testosterone decreased by 70–80%, testicular LH receptors by 26%, those of prolactin by 50% (P<0·01 for all), but those of FSH remained unaffected. Acute (2 h) stimulation by a GnRH agonist (buserelin, 10 μg/kg i.m.) resulted in similar LH, FSH and testosterone responses in the fasted and control animals, and human chorionic gonadotrophin (hCG) stimulation (30 IU/kg i.m.) in similar increases in testosterone. A 42% decrease was found in pituitary content of mRNA of the common α subunit (P<0·05), but the mRNAs of the LH- and FSH-β chains and prolactin were unaffected by fasting for 4 days. When the same mRNAs were measured after 6 days of fasting, the decrease of the mRNA of FSH-β also became significant (50%, P<0·01). In contrast, the mRNA of LH-β was increased twofold (P<0·01) at this time and serum LH levels were similar in control and starved animals. It is concluded that during short-term starvation of male rats: (1) the decrease in gonadotrophin and prolactin synthesis and secretion is first noticed on the level of translation (protein synthesis), and the mRNA levels of these hormones may respond more slowly to starvation, (2) decreased pituitary GnRH receptors indicate decreased GnRH release from the hypothalamus, (3)the gonadotrophin and prolactin loss results secondarily in decreased testicular androgen synthesis and LH and prolactin receptor levels, (4) no decrease occurs during starvation in acute gonadotrophin response to GnRH, or testicular testosterone response to hCG, (5) the primary response to starvation in male rat pituitary-testicular function is the loss of normal hypothalamic support of gonadotrophin and prolactin secretion, rather than direct nutritional effects on the pituitary and testis, and (6) when starvation is continued beyond 4 days, a recovery is seen in pituitary mRNA on the LH-β chain and in serum LH, most probably because the starvation-associated decrease serum testosterone is a more potent positive stimulus of LH synthesis than the direct hypothalamic-pituitary inhibition. Journal of Endocrinology (1989) 121, 409–417


1994 ◽  
Vol 141 (1) ◽  
pp. 143-151 ◽  
Author(s):  
I A Klaij ◽  
M A Timmerman ◽  
P Kramer ◽  
H M A Meijs-Roelofs ◽  
F H de Jong

Abstract Age-related short-term effects of hemicastration on testicular weight, serum FSH, immunoreactive inhibin, LH and testosterone, testicular levels of inhibin subunit mRNA expression, and bioactive and immunoreactive inhibin were studied in rats of 8, 15 and 22 days of age. Hemicastration led to an increased weight of the remaining testis after 24 h in 8- and 15-day-old rats, but not in 22-day-old rats. Serum FSH levels were elevated in all hemicastrated rats after 8 h. However, serum immunoreactive inhibin levels were decreased only after 72 h in 8-day-old rats and after 24 h in 15- and 22-day-old rats. Inhibin α-subunit mRNA expression was increased in the testes of hemicastrated rats of 8 and 15 days of age, whereas inhibin βB-subunit mRNA expression was elevated in the testes of 15-day-old rats but not in those of 8- and 22-day-old rats. The increase in α-subunit mRNA content per testis was caused by an increased concentration and increased testicular weight, whereas the increase in βB-subunit mRNA in the remaining testis parallelled the increased testicular weight, indicating that different mechanisms play a role in the regulation of these mRNAs. In 22-day-old rats, a transiently decreased expression of inhibin βB-subunit mRNA was observed 8 h after hemicastration. The increased inhibin α- and βB-subunit mRNA expression in 8- and 15-day-old rats did not result in increased testicular bioactive and immunoreactive inhibin content of the remaining testis, whereas in 22-dayold rats an increased immunoreactive inhibin content of the remaining testis was observed. These data indicate that efficiency of translation, post-translational modifications or transport from the testis play an important role in determining the final testicular content of inhibin. In conclusion, the response of the remaining testis and the role of inhibin in the regulation of the pituitary-testis axis after unilateral castration depend on the age at which the animals are hemiorchidectomized. Journal of Endocrinology (1994) 141, 143–151


1989 ◽  
Vol 2 (3) ◽  
pp. 213-224 ◽  
Author(s):  
G. Saade ◽  
D. R. London ◽  
M. R. A. Lalloz ◽  
R. N. Clayton

ABSTRACT The effect of castration and gonadal steroid replacement on the concentrations of LH-β and α subunit and prolactin mRNA was examined in mice. Mouse LH-β, α and prolactin mRNAs were approximately 0·8, 0·7 and 1·1 kb in size respectively. After ovariectomy, LH-β mRNA levels increased 2- to 2·5-fold, while α mRNA levels increased 2·5-fold 6 and 10 days after ovariectomy. Serum LH rose after 2 days to reach six times control values at 10 days. Pituitary LH content doubled by 8 days after ovariectomy. Prolactin mRNA levels decreased to 50–60% of control at 3, 6, 8 and 10 days after ovariectomy and parallelled the fall in serum prolaction. Pituitary prolactin content fell more slowly, to 50% of intact control values by 10 days. The increase in both LH-β and α subunit mRNA, and decrease in prolactin mRNA, and serum and pituitary hormone changes, after ovariectomy were prevented by oestradiol or oestradiol plus progesterone replacement. Levels of LH-β mRNA increased more quickly in male than in female mice, theearliest change being seen 24 h after orchidectomy. Maximum values (two- to threefold) were found on day 6 after orchidectomy. Concentrations of α mRNA increased by 12 h to between 2 and 2·5 times control from 3 to 10 days after orchidectomy. Serum LH doubled by 12 h and was three to five times greater than control values up to 10 days. Pituitary LH content fell by 48 h before gradually increasing to intact values after 10 days. Prolactin mRNA levels decreased progressively from 2 days after orchidectomy, and this decrease was preceded by a fall in serum and pituitary prolactin which remained low throughout the experiment. Testosterone treatment attenuated the rise in α mRNA, prevented the rise in LH-β mRNA and serum LH and partially restored the decrease in prolactin mRNA seen after orchidectomy. We conclude that in mice, as in rats and ewes, both LH-β and α subunit mRNAs are negatively regulated by gonadal steroids, whereas prolactin mRNA is positively regulated, although there are temporal differences in patterns of mRNA responses between males and females. By comparison with female rats the rise in LH-β mRNA after ovariectomy was slower in mice. Moreover, the discordant changes in pituitary LH content and LH subunit mRNAs seen in mice after castration were not observed in rats. Furthermore, pituitary prolactin and prolactin mRNA do not fall after orchidectomy of rats. The modest (50%) increase of LH-β mRNA after castration of mice suggests that an increase in mRNA is not necessarily required for increased LH production.


2007 ◽  
Vol 562 (1-2) ◽  
pp. 130-131 ◽  
Author(s):  
Angelika Varga ◽  
Péter Bagossi ◽  
József Tözsér ◽  
Barna Peitl ◽  
Zoltán Szilvássy

1998 ◽  
Vol 85 (5) ◽  
pp. 1903-1908 ◽  
Author(s):  
Ronald R. Gomes ◽  
Frank W. Booth

We examined the age-related association in skeletal muscle between atrophy and expression of mRNAs encoding both the γ-subunit of the nicotinic acetylcholine receptor (AChR), and myogenin, a transcription factor that upregulates expression of the γ-subunit promoter. Gastrocnemius and biceps brachii muscles were collected from young (2-mo-old), adult (18-mo-old), and old (31-mo-old) Fischer 344/Brown Norway F1 generation cross male rats. In the gastrocnemius muscles of old vs. young and adult rats, lower muscle mass was accompanied by significantly elevated AChR γ-subunit and myogenin mRNA levels. In contrast, the biceps brachii muscle exhibited neither atrophy nor as drastic a change in AChR γ-subunit and myogenin mRNA levels with age. Expression of the AChR ε-subunit mRNA did not change with age in either gastrocnemius or biceps brachii muscles. Thus changes in skeletal muscle AChR γ-subunit and myogenin mRNA levels may be more related to atrophy than to chronological age in old rats.


1991 ◽  
Vol 206 (2) ◽  
pp. 165-166 ◽  
Author(s):  
Peter P. Li ◽  
Ying-Kee Pam ◽  
L.Trevor Young ◽  
Jerry J. Warsh

1993 ◽  
Vol 137 (1) ◽  
pp. 69-79 ◽  
Author(s):  
A. Perheentupa ◽  
M. Bergendahl ◽  
F. H. de Jong ◽  
I. Huhtaniemi

ABSTRACT Direct effects of testosterone on gonadotrophins at the pituitary level were studied in intact and castrated immature (age 10 days) and mature (70 days) male rats. Gonadotrophin-releasing hormone action was blocked by treatment with a potent GnRH antagonist, Ac-d-pClPhe-d-pClPhe-d-Trp-Ser-Tyr-d-Arg-Leu-Arg-Pro-d-Ala-NH2CH3COOH (Ant; Organon 30276; 1·0 mg/kg body weight per day) injected subcutaneously. Silicone elastomer capsules were used for the testosterone treatment. Both treatments commenced on the day of orchiectomy and lasted for 7 days. In adult male rats Ant treatment suppressed serum testosterone from 9·5 ± 2·5 (s.e.m.) nmol/l to below the limit of detection (< 0·10 nmol/l; P < 0·01), and the testosterone implants reversed the decrease. Treatment with Ant decreased the pituitary content of FSH-β subunit mRNA in intact and orchiectomized rats to 14% of their respective controls (P < 0·01). These levels were increased to 80–81% of controls (not significant) in both groups by combined treatment with testosterone and Ant. Orchiectomy alone increased FSH-β subunit mRNA by 202% (P < 0·01). In intact immature rats Ant treatment decreased the level of pituitary FSH-β subunit mRNA to 21% (P<0·01), and a partial recovery (P < 0·01) to 42% of controls was observed with combined Ant + testosterone treatment. In contrast, in orchiectomized immature rats, where ANT decreased FSH-β subunit levels to 48% of controls (P < 0·01), testosterone was able to reverse these mRNA levels completely (114% of controls). No evidence for the direct pituitary effects of testosterone were found in the mRNA of the common α or LH-β subunits. In adult rats, the testicular inhibin α and βA subunit mRNA levels were increased (P < 0·01) by Ant + testosterone compared with Ant-treated animals, but there were no differences in serum immunoreactive inhibin between any of the uncastrated adult groups. In intact immature rats, Ant + testosterone treatment increased (P < 0·01) inhibin βA subunit mRNA levels compared with controls and Ant-treated animals. Ant decreased the level of peripheral inhibin immunoreactivity from 8·3 ± 2·0 U/ml to 2·1 ± 0·4 U/ml (P < 0·01) and testosterone reversed it to 5·8 ± 0·6 U/ml (not significant). In conclusion, our observations indicated that testosterone is able to stimulate FSH gene expression and secretion directly in immature and adult rats, but the testosterone response is enhanced at both ages by orchiectomy, even more so in the immature rat. This may be explained by age differences in the contribution of testicular inhibin to the regulation of FSH synthesis and secretion at the pituitary level. Journal of Endocrinology (1993) 137, 69–79


1994 ◽  
Vol 143 (2) ◽  
pp. 353-358 ◽  
Author(s):  
O Lapcik ◽  
A Perheentupa ◽  
M Bicikova ◽  
I Huhtaniemi ◽  
R Hampl ◽  
...  

Abstract The effects of 3-week treatment with increasing doses of epitestosterone (ET) on gonadotrophin gene expression and secretion, on testosterone and 5α-dihydrotestosterone (DHT) levels, and on the weight of testes and prostates, were studied in intact adult male rats. The hormones were delivered by means of silastic capsules of different lengths filled with the steroid. One group of rats received testosterone (T) instead of ET, to compare the results with previous studies concerning the testosterone effect. The controls were given capsules with glucose only. Treatment with ET, as well as with T, significantly reduced the weights of prostates. When the data from ET-treated rats and controls were combined, a significant negative correlation (P<0·001) was found between the weight of prostates and serum ET. T, in contrast to ET, also decreased significantly the weights of testes. ET treatment caused a significant reduction of serum T levels but only an insignificant decline of DHT levels, independent of the dose. Serum and pituitary (p) luteinizing hormone (LH) levels in the ET-treated rats did not change. Pituitary mRNA contents for the βLH subunit (βLH-mRNA) showed a dose-dependent significant increase, up to 170% (P<0·01), with ET treatment. pFSH decreased with the lowest ET (2 cm) dose (P<0·05), but no change was observed with the other doses. The mRNA for the common α-subunit also increased with the ET load. In conclusion, ET acts at several sites in the regulation of gonadotrophin formation and release. It enhances the steady-state mRNA levels of both gonadotrophins in the pituitary. At the same time, ET may act directly in the pituitary by inhibition of post-transcriptional events in LH synthesis. A direct inhibitory effect of ET at the hypothalamic level is also possible. The circulating levels of both gonadotrophins are thus the result of these composite effects. Journal of Endocrinology (1994) 143, 353–358


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