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Analysis of the primary spermatocytes during spermatogenesis in the mouse.

小鼠精子发生过程中初级精母细胞的分析。

基本信息

批准号：
06670009
负责人：
MAEKAWA Mamiko
金额：
$ 1.28万
依托单位：
Chiba University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1994
资助国家：
日本
起止时间：
1994 至 1995
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-06670009/
关键词：
Spermatogenesis Testis Spermatocyte Monoclonal antibody Actin filament 減数分裂

项目摘要

(1) Monoclonal antibodies were raised against primary spermatocytes. Using indirect immunofluorescence assay and immunoelectron microscopy, it has confirmed that three antibodies recognize surface antigens of the different stages of spermatocytes in the mouse testis. Antibody 10B3 recognizes spermatocytes during middle to late pachytene stages (VI-X) , antibody 10D6 stains spermatocyte during leptotene to early pachytene stages (I-IV) and antibody 12F7 labeled spermatocytes during late stages(VII-VIII).(2) Antibodies 10D6 and 12F7 also recognized specific sites of the surface of the sperm head. To see the effects of the antibodies on the fertilization, the antibodies were added to the culture medium of the in vitro fertilization system, but obvious effects were not obtained.(3) Antigens recognized by the three antibodies were characterized by Western blotting. Antibodies 10D6 and 12F7 failed to detect any spot, whereas antibody 10B3 labeled one spot (MW 50kD,pI5.5) by two-dimensional Western blotting.(4) We tried to culture spermatogenic cells including spermatocytes and gonocytes. However, significant results were not obtained.(5) Spermatocytes move from the basal to adluminal compartment in the seminiferous tubule during maturation, and cytoskeletal proteins such as actin filaments might influence the movement of the spermatocytes. Thus we investigated the distribution of actin filaments and actin-binding proteins in the testis. As a result, the distribution of actin filaments changed during postnatal development. Furthermore, in the cryptorchid testis, in which spermatogenesis was disturbad, the arrangement of actin filaments was also disrupted. These results suggest the relation between the distribution of the actin filaments and spermatogenic activity.

(1)针对初级精母细胞产生单克隆抗体。利用间接免疫荧光法和免疫电镜技术，证实三种抗体能够识别小鼠睾丸中不同阶段精母细胞的表面抗原。抗体 10B3 识别粗线期中后期 (VI-X) 的精母细胞，抗体 10D6 在细线期到粗线期早期 (I-IV) 期间对精母细胞进行染色，抗体 12F7 标记晚期 (VII-VIII) 的精母细胞。 (2) 抗体 10D6 和 12F7 还识别精母细胞的特定位点。精子头部的表面。为了观察抗体对受精的影响，将抗体添加到体外受精系统的培养基中，但没有获得明显的效果。(3)通过Western blotting对三种抗体识别的抗原进行表征。通过二维Western blotting，抗体10D6和12F7未检测到任何斑点，而抗体10B3标记了一个斑点(MW 50kD，pI5.5)。(4)我们尝试培养生精细胞，包括精母细胞和性母细胞。然而，并没有获得有意义的结果。(5)精母细胞在成熟过程中从曲细精管的基底室移动到腔室，肌动蛋白丝等细胞骨架蛋白可能会影响精母细胞的运动。因此，我们研究了肌动蛋白丝和肌动蛋白结合蛋白在睾丸中的分布。结果，肌动蛋白丝的分布在出生后发育过程中发生了变化。此外，在隐睾中，精子发生受到干扰，肌动蛋白丝的排列也被破坏。这些结果表明肌动蛋白丝的分布与生精活性之间的关系。