Molecular Biological Studies of the Mechanisms in Amphibian Spermatogenesis.

两栖动物精子发生机制的分子生物学研究。

基本信息

批准号：
01480029
负责人：
ABE Shin-ichi
金额：
$ 4.35万
依托单位：
Kumamoto University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (B)
财政年份：
1989
资助国家：
日本
起止时间：
1989 至 1991
项目状态：
已结题

项目摘要

I. Differentiation from secondary spermatogonia to primary spermatocytes by FSH in organ culture of testes fragments from the newt. Cynops pyrrhogaster : When testes fragments rich in secondary spermatogonia were cultured in the presence of follicle-stimulating hormone (FSH) alone, primary spermatocytes appeared in more than half of the cysts by the second week. These results indicate that FSH is indispensable and sufficient for the differentiation from spermatogonia to primary spermatocytes.II. How is the flagellar length of mature sperm determined ? Comparison of flagellar growth in round spermatids between Cynops and Xenopus : The flagellar length of Cynops sperm is 13 times longer than that of Xenopus sperm. The rate and period of flagellar growth in Cynops spermatids were 2 to 3-fold those in Xenopus. The period of flagellar elongation almost coincided with that of tubulin synthesis. The differences in the rate and period of tubulin synthesis between Cynops and Xenopus seems to be … More due to the difference in the stability of tubulin mRNAs between the two species.III. Synthesis of sperm-specific basic nuclear proteins (SPs) in spermatids and expression of SP4 mRNA in primary spermatocytes from Xenopus laevis : In synchronous culture of round spermatids, the synthesis of SP1, 3-5 began by day 3. CDNA clones for SP4 and 5 were isolated from a cDNA library from round spermatids. The Northern blots showed that SP4 and 5 mRNAs were present in primary spermatocytes as well as in round spermatids. This result shows that SP4 and 5 genes are transcribed in the primary spermatocyte stage but are translated at the round spermatid stage.IV. Role of manchette (microtubule bundles) in nuclear elongation in spermatids from Cynops pyrrhogaster : The role of microtubules in nuclear shaping of spermatids has yet to be determined. We found that manchette structure in newt elongate spermatids is resistant to the treatment of Ca^<2+> and low temperature. 1M NaCl Sup extracted from low temp-stable microtubules in testes promoted polymerization of tubulin at 37 C and resisted depolymerization at 0 C in a dose-dependent manner. These results indicate that 1M NaCl Sup contains MAPs (microtubule associated proteins) which promote tubulin polymerization and confer the resistance against tubulin depolymerization. Less

I.从纽特的测试片段中，FSH从二级精子症到原代精子细胞的分化。 Cynops pyrrhogaster：仅在仅在诱导叶片的马龙（FSH）的情况下培养富含继发精子的测试片段时，第二周到第二周，一半以上的囊肿出现了原代精子细胞。这些结果表明，FSH是必不可少的，并且足以分化从精子症到原代精子细胞。II。如何确定成熟精子的旗杆长度？比较cy和爪蟾之间圆形精子中的旗杆生长：cynop的旗杆长度比爪蟾精子长13倍。 cynop的精子中旗杆生长的速率和周期为2至3倍。鞭毛伸长的时期几乎与微管蛋白合成的时期相吻合。细节和爪蟾之间的小节蛋白合成速率和周期的差异似乎是……更多是由于两种物种之间小管蛋白mRNA的稳定性差异。精子中精子特异性核蛋白（SP）的合成和SP4 mRNA的表达和Sp4 mRNA的表达来自Xenopus laevis的原代精子细胞：在圆形精子的同步培养中，Sp1，3-5的SP1，3-5的合成开始，从Sp4和5。北印迹表明，原代精子细胞以及圆形精子中存在SP4和5个mRNA。该结果表明，SP4和5基因在原发性精子阶段转录，但在精子阶段进行了翻译。 Manchette（微管束）在Cynops Pyrrhogaster的精子中的作用：微管在精子核成型中的作用尚未确定。我们发现Newt伸长式精子中的Manchette结构可抵抗Ca^<2+>和低温的治疗。在测试中从低温稳定的微管中提取的1M NaCl SUP促进了微管蛋白在37 C下的聚合，并以剂量依赖性方式在0 C下抗性。这些结果表明，1M NaCl SUP包含图（微管相关蛋白），这些图（促进小节蛋白）聚合和会议对小管蛋白沉积的抗性。较少的