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Comparative studies between in vivo and in vitro spermatogenesis of Japanese eel (Anguilla japonica)

机译：日本鳗（Anguilla japonica）体内和体外精子发生的比较研究

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In order to check the quality of in vitro spermatogenesis of Japanese eel, in vitro11-ketotestosterone (11-KT) induced spermatogenesis was compared with in vivo spermatogenesis induced by a single injection of human chorionic gonadotropin (hCG) in detail. DNA contents of germ cells from in vitro and in vivo testicular fragments were compared using flow cytometry Since the in vitro result of flow cytometry showed prominent 1C peak including spermatozoa and spermatids, the reduction of DNA by meiosis was assumed to progress normally, (i.e., haploid spermatozoa were produced in this in vitro system). In the testes of in vitro culture, however, spermatozoa were not released into lumen. Furthermore, the number of mitotic divisions of the in vitro experiment (6 divisions) was fewer than that of in vivo (10 divisions). In electron microscopy observations, both of in vivo and in vitro spermatozoon had a crescent-shaped nucleus with a flagellum, and a single large spherical mitochondrion. However, the elongation of the sperm head was not sufficient and the mitochondrion was not always located at the anterior end as is observed for the spermatozoa obtained from hCG injected eels. Eel spermatogenesis related substance-11 (eSRS11) is homologue of histone Hill which is up-regulated during spermatogenesis. Using this probe, in vitro spermatogenesis was also evaluated in molecular levels. In Northern blot analysis, eSRS11 mRNA was detected in both in vivo and in vitro testes. However, the expression of in vitro was much weaker than that of in vivo. These differences indicate that the stimulation of 11-KT is not sufficient, and another factors are needed to induce complete spermatogenesis in vitro.
机译：为了检查日本鳗鱼体外精子发生的质量，将体外11-酮睾酮（11-KT）诱导的精子发生与单次注射人绒毛膜促性腺激素（hCG）诱导的体内精子发生进行了比较。使用流式细胞术比较了体外和体内睾丸碎片中生殖细胞的DNA含量。由于流式细胞术的体外结果显示包括精子和精子在内的1C峰很显着，因此减数分裂导致的DNA还原正常进行，（即，在该体外系统中产生了单倍体精子。然而，在体外培养的睾丸中，精子没有释放到管腔中。此外，体外实验的有丝分裂分裂数（6个分裂）少于体内的有丝分裂分裂的数目（10个分裂）。在电子显微镜观察中，体内和体外的精子均具有带鞭毛的新月形核和单个大的球形线粒体。然而，如从hCG注射的鳗鱼中获得的精子所观察到的那样，精子头的伸长是不够的，并且线粒体并不总是位于前端。鳗鱼精子生成相关物质11（eSRS11）是在精子发生过程中上调的组蛋白Hill的同源物。使用该探针，还可以在分子水平上评估体外精子发生。在RNA印迹分析中，在体内和体外睾丸中都检测到了eSRS11 mRNA。但是，体外表达远弱于体内表达。这些差异表明11-KT的刺激是不够的，还需要另一个因素来诱导体外完全精子发生。

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