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Establishment of in vitro culture system of seminiferous epitherium and clarifying the mechanism of spermatogenesis

生精上皮体外培养体系的建立及精子发生机制的阐明

基本信息

批准号：
02454098
负责人：
HAYASHI Yoshihiro
金额：
$ 3.46万
依托单位：
The University of Tokyo
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (B)
财政年份：
1990
资助国家：
日本
起止时间：
1990 至 1991
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-02454098/
关键词：
seminiferous eitherium in vitro culture system mouse testis spermatic cord Sertoli cells spermatogenic cells sexual differentiation glycoprotein

项目摘要

To establish the in vitro culture system of the seminiferous epitherium is an important research subject for clarifying the mechanisms of both the sexual differentiation and the spermatogenesis. In the present study, results were conipbsed of three parts, and all of them were published in 8scientific papers. First, we demonstrated the stage-specific appearance of glycoproteins and glycolipeds on the cell membrane of sertoli cells and germ cells in mouse testes. Using in vitro culture system, the role of these molecules was clarified. by adding the inhibitors against correspondinq moldcules, that is, the strong involvement of these molecules in the spermatogenesis. Second, to find outthe essential factors on spermatogenesis, morphological-characteristics of interstitial tissues was examined in the boar testis. The separation mechanism between the basal and abluminal compartments in the seminiferous epitherium was first indicated by observing the cytoskeleton in the ectoplasmic specialization (bloodtestis barrier) in boar testis by deep-eecth method. Further, monoclonal antibodies specific to-germ cells were cloned out in this study, ana a 103 KDa protein was detected in the cells from pachytene spermatocytes to early spermatids. Third, to clarify the counter-current transfer of substances from the testicular vein to the artery was morphologically examined using three kinds of mammals, and both the common and specific architectures were demonstrated in this study.

建立生精上皮细胞的体外培养体系是阐明生精上皮细胞性分化和精子发生机制的重要研究课题。本研究共分三个部分，共发表了80篇论文。首先，我们证明了小鼠睾丸支持细胞和生殖细胞细胞膜上的糖蛋白和糖脂的阶段特异性外观。利用体外培养系统，阐明了这些分子的作用。通过加入抑制相应的分子的抑制剂，即这些分子在精子发生中的强烈参与。第二，研究了猪睾丸间质组织的形态学特征，以探讨影响精子发生的重要因素。通过对猪精巢外质特化（血睾屏障）细胞骨架的观察，首次揭示了生精上皮基腔室和近腔室的分离机制。本研究还克隆了生殖细胞特异性单克隆抗体，并在粗线期精母细胞至早期精子细胞中检测到一个分子量为103 KDa的蛋白。第三，为了阐明物质从睾丸静脉到动脉的逆流转移，本研究使用三种哺乳动物进行了形态学检查，并在本研究中展示了共同和特定的结构。