MOLECULAR ANALYSIS OF HUMAN SPERM ACROSOMAL MOLECULE H-SP-1 IN SPERM-EGG INTERACTION

人精子顶体分子 H-SP-1 在精卵相互作用中的分子分析

• 批准号: 09671723
• 负责人: KOMORI Shinji
• 金额: $ 1.86万
• 依托单位: HYOGO COLLEGE OF MEDICINE
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1998
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09671723/
• 关键词: sperm-egg interaction; acrosome; human; membrane molecule; calcium channel; 精子失体膜分子

Several surface molecules of both gametes are involved in gametes recognition during fertilization( Saling, 1989 ). These molecules are responsible for each process of fertilization and species-specific interaction between the gametes. Studies of these molecules may be essential for understanding the mechanism of sperm-egg interaction and for the development of a contraceptive vaccine.We isolated a human sperm antigen gene (h-Sp-l) by screening from human testis cDNA library with human sperm specific antisera. The sequence analysis of the h-Sp-l gene indicated that it contained 2170 bp and had 45% homology to human synaptophysin gene, which was considered to play a crucial role in calcium dependent signal transduction in synaptic vesicles in the terminal end of nerves. The analysis of the deduced amino acids from the h-Sp- 1 gene showed that it encoded 259 amino acids containing four hydrophilic regions and four hydrophobic regions. This indicated that the molecules encoded by the h-Sp- 1 gene might be a transmembrane protein with four membrane spanning regions like the synaptophysin. Northern blot analysis indicated that h-Sp-1 gene had 2 kind of transcripts ( 2.3 kb and 1.1kb ). 2.3kb transcript expressed ubiquitously whereas 1.1kb transcript expressed abundantly in human testis. In order to characterize the molecules encoded by the h-Sp-l gene, the antisera were generated by immunizing rabbits with synthetic polypeptides ( aminoacid No 174-198 ) based on the h-Sp-1 gene. Western blot analysis indicated that the antisera reacted a 40 kDa molecule. The antisera also stained fixed human sperm head by indirect immunofluorescence staining. The purified antisera showed the inhibitory effect on human sperm penetration assay into zona-free hamster eggs. However, they did not show any sperm immobilizing and agglutinating activities. These results suggested that the molecules encoded by the h-Sp-1 gene might play an important role in the sperm-egg interaction.

两个配子的几个表面分子参与受精过程中配子的识别（Saling, 1989）。这些分子负责每个受精过程和配子之间的物种特异性相互作用。对这些分子的研究可能对了解精子-卵子相互作用的机制和开发避孕疫苗至关重要。用人精子特异性抗血清从人睾丸cDNA文库中筛选分离出一个人精子抗原基因h- sp - 1。h- sp - 1基因序列分析表明，其长度为2170 bp，与人突触素基因同源性为45%，被认为在神经末梢突触囊泡钙依赖信号转导中起重要作用。从h-Sp- 1基因推导出的氨基酸序列分析表明，该基因共编码259个氨基酸，其中包含4个亲疏水区和4个疏水区。这表明h-Sp- 1基因编码的分子可能是一种与synaptophysin一样具有四个跨膜区的跨膜蛋白。Northern blot分析表明，h-Sp-1基因有2种转录本（2.3 kb和1.1kb）。2.3kb的转录本在人睾丸中普遍表达，而1.1kb的转录本在人睾丸中大量表达。为了鉴定h-Sp-1基因编码的分子，用合成的h-Sp-1基因多肽（氨基酸No 174-198）免疫家兔制备了抗血清。Western blot分析表明，该抗血清反应分子量为40kda。用间接免疫荧光法对固定的人精子头进行染色。纯化后的抗血清对人精子穿透无带仓鼠卵有抑制作用。然而，它们没有表现出任何精子的固定和凝集活性。这些结果表明，由h-Sp-1基因编码的分子可能在精子-卵子相互作用中起重要作用。

项目成果

Shinji Komori, Kazuko Sakata, Hiroyuki Tanaka, Ri-ichiro Kanazawa, Yoshiyuki Tsuji, Koji Koyama and Shinzo Isojima.: "Synaptophysin related molecule in human sperm : the role in sperm-egg interaction" (submitted).

Shinji Komori、Kazuko Sakata、Hiroyuki Tanaka、Ri-ichiro Kanazawa、Yoshiyuki Tsuji、Koji Koyama 和 Shinzo Isojima.：“人类精子中的突触素相关分子：在精卵相互作用中的作用”（已提交）。