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通过组织切片和电子显微镜观察经多次注射人绒毛膜促性腺激素(human chorionic gonadotropin,hCG)和鲤鱼脑垂体匀浆液(carp pituitary extract,CPE)促熟的雄性鳗鲡(Anguilla japonica)精巢和脑垂体超微结构的变化,并采用超高效液相色谱-串联质谱(UPLC/MS/MS)联用的方法,观察性腺和血液中7种类固醇激素含量的变化。结果发现:成熟组鳗鲡精巢发育至第Ⅴ期,排精10 d时鳗鲡精巢进入退化吸收期。性成熟雄性鳗鲡的脑垂体GtH分泌细胞代谢活动旺盛,细胞中分泌小球增多,囊泡数量增加,线粒体和内质网丰富。鳗鲡精巢和血液中睾酮(testosterone,T)、17α,20β-二羟基-4-孕烯-3-酮(17α,20β-dihydroxy-4-pregnen-3-one,DHP)和17α-羟孕酮(17α-hydroxyprogesterone,17α-OHP)3种激素含量较高,且成熟的和排精10 d后雄鱼精巢和血液中T、DHP和17α-OHP的含量均显著高于对照组(P﹤0.05),排精10 d时雄鱼精巢和血液中17α-羟孕酮的含量显著低于成熟组鳗鲡精巢和血液中的含量(P﹤0.05);精巢和血液中未检出孕酮(progesterone,P)、雌酮(estrone,E1)、β-雌二醇(estradiol-17β,E2)和雌三醇(estriol,E3)4种激素。结果表明,睾酮、17α-羟孕酮和DHP是雄鱼成熟的关键类固醇激素,其含量在性成熟时达到峰值,排精10 d时含量降低,其含量受脑垂体GtH分泌的调控。
The testis and pituitary gland of male Anguilla japonica which were induced by multiple injections of human chorionic gonadotropin (hCG) and carp pituitary extract (CPE) were observed by histological sections and electron microscopy. Ultrastructural changes, and using ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC / MS / MS) combined method to observe the changes of gonadal and blood 7 kinds of steroid hormones. The results showed that in the mature group, the testis of Anguilla anguilla developed to the stage Ⅴ, and the testis of Anguilla anguillarum entered the stage of degenerative absorption after 10 days of ejaculation. GtH secretion in pituitary gland of sexually mature male Anguilla anguillarum had a strong metabolic activity, increased secretion of small cells in the cells, increased number of vesicles and abundant mitochondria and endoplasmic reticulum. Testosterone (T), 17α, 20β-dihydroxy-4-pregnen-3-one (DHP) and 17α-hydroxyprogesterone (17α-hydroxyprogesterone, 17α-OHP), and the content of T, DHP and 17α-OHP in testis and blood of mature fish and blood 10 d after sperm maturation and 10 days of sperm injection were significantly higher than those of control group (P <0.05 ), The content of 17α-hydroxyprogesterone in the testis and blood of male fish during 10 days of ejaculation was significantly lower than that of the mature group (P <0.05), while no progesterone was detected in the testis and blood P, estrone (E1), estradiol-17β (E2) and estriol (E3). The results showed that testosterone, 17α-hydroxyprogesterone and DHP were the key steroid hormones in males. The content reached the peak at sexual maturity. The content decreased after 10 days of ejaculation and was regulated by the secretion of GtH in pituitary gland.