EXPRESSION OF HEAT-SHOCK GENES IN MOUSE SPERMATOGENIC CELLS

热休克基因在小鼠生精细胞中的表达

• 批准号: 3855971
• 负责人: E M EDDY
• 金额: --
• 依托单位: NATIONAL INSTITUTE OF ENVIRONMENTAL HEALTH SCIENCES
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3855971
• 关键词: affinity chromatography; antiserum; complementary DNA; environmental stressor; gene expression; genetic library; genetically modified animals; germ cells; heat; laboratory mouse; meiosis; regulatory gene; sperm; spermatogenesis; stress proteins; testis; western blottings

The synthesis of heat-shock proteins (hsp) following environmental stress is a highly conserved response and apparently protects cells against long-term effects of adverse conditions. One of the most abundant proteins (P70) in spermatogenic cells is related closely to hsp7O, as shown by ATP affinity chromatography purification, immunoblots of two-dimensional protein gels, and peptide mapping. The P70 protein is synthesized during the meiotic phase of spermatogenesis. Although several members of the hsp7O gene family have been sequenced and their mRNA expression characterized, little is known about the proteins encoded by these genes. We developed antisera specific to P70 and hsp7O to investigate the expression and role of these proteins in male germ cells. The antiserum to P70 was also used to select clones from a mouse spermatocyte cDNA library. One of these clones hybridized to a testis-specific, developmentally regulated 3.0 kb mRNA whose expression was not affected by heat. Sequence analysis indicated that the clone has high homology to previously reported genes of the hsp70 family. However, it appears that spermatogenic cells do not produce hsp70 mRNA or protein in response to heat stress. This suggests that P70 gene expression is linked to suppression of the hsp7O gene in spermatogenic cells. However, the process of spermatogenesis is unusually sensitive to elevation of temperature and to many toxic agents and P70 may function in germ cell differentiation rather than to protect against the effects of environmental stress. Although P70 expression may be beneficial to the process of spermatogenesis, this process may also leave male germ cells particularly susceptible to environmental stress. Genomic clones have been isolated for the P70 gene and transgenic mice will be produced to examine the spermatogenic cell-specific regulation of expression of this gene.

环境胁迫后热休克蛋白（HSP）的合成 是一种高度保守的反应，显然可以保护细胞免受 不利条件的长期影响。 最丰富的蛋白质之一 (P70)生精细胞内的ATP含量与hsp7O密切相关 亲和层析纯化，二维免疫印迹 蛋白质凝胶和肽图谱。 P70蛋白是在 精子发生的减数分裂阶段。 虽然几个成员的 hsp70基因家族已被测序，其mRNA表达 虽然这些基因的特征是，但对这些基因编码的蛋白质知之甚少。 我们研制了P70和hsp70的特异性抗血清， 这些蛋白在雄性生殖细胞中的表达和作用。 抗血清 P70也被用于从小鼠精母细胞cDNA文库中选择克隆。 其中一个克隆与睾丸特异性的，发育性的 调节的3.0kb mRNA的表达不受热的影响。 序列 分析表明，该克隆与已报道的同源性较高 HSP70家族的基因 然而，生精细胞似乎 不产生热休克蛋白70的mRNA或蛋白。 这 提示P70基因表达与hsp7O抑制有关， 基因在生精细胞中的表达。 然而，精子发生的过程是 对温度升高和许多有毒物质异常敏感 P70可能在生殖细胞分化中起作用，而不是保护 对抗环境压力的影响。 虽然P70表达可能 有利于精子发生的过程，这个过程也可能 使雄性生殖细胞特别容易受到环境压力影响。 已经分离出P70基因的基因组克隆， 以检查生精细胞特异性调节 这个基因的表达。