Pubertal development of male African catfish,Clarias gariepinus. In vitro steroidogenesis by testis and interrenal tissue and plasma levels of sexual steroids

In fish, sex steroids initiate and/or accelerate the maturation of the brain-pituitary-gonad axis. In order to obtain information on the steroid milieu during the pubertal development of male African catfish, we have monitored the conversion of [³H]-pregnenolone and [³H]-androstenedione by testis and [³H]-pregnenolone by interrenal tissue fragmentsin vitro. Pubertal development occurs between two and six months of age. Testicular development proceeds through four main stages that are characterised histologically by the presence of spermatogonia (stage I), spermatogonia and spermatocytes (stage II), spermatogonia, spermatocytes and spermatids (stage III), and all germ cells including spermatozoa (stage IV). 11β-Hydroxyandrostenedione and cortisol were the main products of testes and interrenal tissue, respectively, in all stages of the pubertal development; a change in the steroidogenic pattern was not observed during this period. The interrenal tissue displayed no significant conversion of [³H]-pregnenolone to 11-oxygenated androgens. Blood plasma was analyzed for the presence of five androgens; testosterone, 11β-hydroxytestosterone, 11β-hydroxyandrostenedione, androstenetrione, and 11-ketotestosterone. 11-Ketotestosterone was the quantitatively dominating androgen in the circulation at all stages of development, which was more pronounced in stages III and IV. The obvious differences between thein vitro andin vivo results, namely 11β-hydroxyandrostenedione being the main testicular productvs. 11-ketotestosterone dominating in the blood, may indicate that 11β-hydroxyandrostenedione is converted to 11-ketotestosterone at extratesticular sites.     

In vitro synthesis of 20α-reduced and of 11- and 21-oxygenated steroids and their sulfates by testes of the goldfish (Carassius auratus): Testicular synthesis of corticosteroids

Testes from spermiating goldfish were incubated with [³H]17-hydroxyprogesterone. The major products in the unconjugated fraction were identified as androstenedione, androstenetrione, 11-β-hydroxyandrostenedione, 11-ketotestosterone, 17,20α-dihydroxy-4-pregnen-3-one (17,20αP) and 11-deoxycortisol. Testosterone was present predominantly in the glucuronide fraction, but yields were low (1–3%). The major components of the sulfate fraction were 17,20αP and 11-deoxycortisol. The identification of cortisone in low yield (< 2.5010) in both the free and sulfate fractions is the first report of corticosteroid biosynthesis by a teleost testis. The high yields of 17,20αP and 11-deoxycortisol and their sulphates suggests that their possible role in spermiation of the goldfish should be further investigated.     

Extrapituitary gonadotropin-releasing hormone (GnRH) binding sites in goldfish

In teleosts, as in other vertebrates, the secretion of pituitary gonadotropin (GTH) is mediated by the hypothalamic decapeptide, gonadotropin-releasing hormone (GnRH). Recent findings in teleosts indicate that GnRH receptors are not restricted to the pituitary gonadotropes and are also associated with somatotropes as well as being present in a number of other tissues. In the present study, we provide novel information on GnRH binding in a number of extrapituitary tissues in goldfish. However, we do not intend to provide full characterization of GnRH binding sites in various extrapituitary tissues in goldfish as this would clearly be outside the scope of this paper. In this study we examined GnRH binding in a number of extrapituitary tissues in goldfish and observed specific binding in ovary, testis, brain, liver and kidney. No specific GnRH binding was observed in muscle, skin, gut, gill and heart. In general, the present findings together with the results of other studies carried out in our laboratory demonstrate that mature goldfish ovary and testis contain two classes of GnRH binding sites, high affinity/low capacity and low affinity/high capacity sites with binding characteristics similar to those of the pituitary GnRH receptors. The brain of goldfish was also found to contain two classes of GnRH binding sites, a super-high affinity/low capacity and a low affinity/high capacity sites. Furthermore, study of goldfish liver and kidney demonstrated the presence of a single class of GnRH binding sites with characteristics different from those of pituitary, ovary, testis and brain. Overall, it is evident that goldfish contains a family of GnRH binding sites which can be classified into four groups based on binding affinities: 1) A class of high affinity binding sites present in the pituitary, ovary and testis, 2) a class of super high affinity sites so far only detected in the brain, 3) a class of intermediate-affinity GnRH binding sites in the liver and kidney, and 4) a class of low affinity binding sites present in all the tissues containing specific GnRH binding sites except for liver and kidney.     

In vitro steroidogenesis by gonads of the Russian sturgeon, Acipenser gueldenstaedti Brandt

Ovaries and testes from the Russian sturgeon at different stages of sexual maturity were incubated with ;sup3;H-androstenedione or ³H-pregnenolone. The major metabolites of androstenedione in both sexes were testosterone and 5- and 5ß-androstanediols, but no evidence was found for the gonadal production of 11-oxygenated androgens. Pregnenolone was converted to 17-hydroxyprogesterone, androstenedione and testosterone together with reduced metabolites. 11-Oxygenated androgens were found in female serum by gas chromatography-mass spectrometry (GC-MS), in serum of both sexes by radioimmunoassay (RIA), and was detectable by RIA in interrenal but not gonadal tissue. The results suggest that sturgeon may differ from teleosts in that 11-oxygenation may take place in extragonadal tissues.     

氟对不同日龄雄性大鼠睾丸组织中表皮生长因子及其受体表达的影响

为探讨氟对不同日龄雄性Wistar大鼠睾丸组织中表皮生长因子（EGF）及其受体（EGFR）表达的影响,选用40日龄雄性Wistar大鼠200只,随机分为2组分别饮用含150mg/L氟化钠的去离子水和去离子水,并于50,80,100,120日龄时处死大鼠,对睾丸组织中EGF及其EGFR的表达情况进行分析。结果表明,染氟组50日龄时睾丸组织间质细胞、精原细胞和生精细胞中EGF的表达量极显著下降（P〈0.01）,精子细胞和管腔脱落物中EGF的表达量降低不明显（P〉0.05）;80,100,120日龄时虽有下降趋势,但无统计学意义（P〉0.05）。而染氟组睾丸组织中EGFR的表达量在50,80,100日龄时均降低,但50,80日龄睾丸组织精子细胞和100日龄睾丸组织曲细精管管腔脱落物中EGFR的表达量极显著下降（P〈0.01）,50日龄时生精细胞中EGFR的表达量显著降低（P〈0.05）;而120日龄时睾丸组织中EGFR的表达量均增高,且管腔脱落物中极显著增高（P〈0.01）。证明氟可降低雄性Wist-ar大鼠性成熟时EGF的表达量以影响精子的生长发育,长期抑制EGF的表达使EGFR的表达增高影响生殖功能。