Yu J Y, Lin T M
Gen Comp Endocrinol. 1986 Jun;62(3):428-36. doi: 10.1016/0016-6480(86)90053-5.
The objectives of this study were to establish a suitable and validated in vitro bioassay of piscine gonadotropins (GTHs) by using a carp testis androgen production system and to compare the androgenic responses in such an assay to gonadotropins from various vertebrate species. The testes from mature carp with gonadosomatic indices of 8-30% were used. Androgen production was first compared with respect to methods for preparation of the carp testis (sliced, minced, homogenized, and collagenase-dispersed testis preparations). The time course of androgen formation, the effects of xanthine and theophylline, and other factors on androgen production also were investigated. Theophylline was more effective than xanthine in potentiation of gonadotropin-evoked androgen formation by carp testis. The testis preparations were incubated in medium 199 (pH 7.40) containing 2 mM theophylline with shaking at 100 cycles/min at 25 degrees C for 4 hr. Homogenized testis preparations had limited ability for androgen production, while sliced, minced, and minced-collagenase-dispersed testis preparations were highly responsive to gonadotropins for androgen production. The minced testis preparation, utilizing 100 mg/ml incubation medium per vial, was chosen as the standard incubation procedure in this study. The minced testis androgen production assay was highly sensitive to gonadotropins from several piscine species (silver carp, common carp, and salmon), and all these GTHs produced parallel dose-related androgen production curves. Mammalian GTHs were also capable of promoting androgen formation by carp testis, but they were much less potent than were piscine GTHs. Pregnant mares' serum gonadotropin (PMSG) was more effective than human chorionic gonadotropin (hCG) in evoking carp testis androgen production.(ABSTRACT TRUNCATED AT 250 WORDS)
本研究的目的是利用鲤鱼睾丸雄激素生成系统建立一种合适且经过验证的鱼类促性腺激素(GTHs)体外生物测定法,并比较该测定法中不同脊椎动物物种促性腺激素的雄激素反应。使用性腺指数为8 - 30%的成熟鲤鱼的睾丸。首先比较了鲤鱼睾丸不同制备方法(切片、切碎、匀浆和胶原酶分散的睾丸制剂)对雄激素生成的影响。还研究了雄激素形成的时间进程、黄嘌呤和茶碱的作用以及其他影响雄激素生成的因素。在增强鲤鱼睾丸促性腺激素诱发的雄激素形成方面,茶碱比黄嘌呤更有效。将睾丸制剂在含有2 mM茶碱的199培养基(pH 7.40)中于25℃、100次/分钟振荡孵育4小时。匀浆的睾丸制剂雄激素生成能力有限,而切片、切碎以及切碎 - 胶原酶分散的睾丸制剂对促性腺激素刺激雄激素生成反应强烈。本研究选择每瓶使用100 mg/ml孵育培养基的切碎睾丸制剂作为标准孵育程序。切碎睾丸雄激素生成测定法对几种鱼类(鲢鱼、鲤鱼和鲑鱼)的促性腺激素高度敏感,所有这些GTHs都产生了平行的剂量相关雄激素生成曲线。哺乳动物的GTHs也能够促进鲤鱼睾丸雄激素的形成,但效力远低于鱼类GTHs。孕马血清促性腺激素(PMSG)在诱发鲤鱼睾丸雄激素生成方面比人绒毛膜促性腺激素(hCG)更有效。(摘要截短于250字)