EXPRESSION OF HEAT-SHOCK GENES IN MOUSE SPERMATOGENIC CELLS

环境应激后热休克蛋白的合成 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)在生精细胞中与hsp70密切相关。 亲和层析纯化双向免疫印迹 蛋白质凝胶和多肽图谱。P70蛋白的合成是在 精子发生的减数分裂阶段。尽管该委员会的几名成员 Hsp70基因家族已测序及其mRNA表达 由于这些基因的特征，人们对这些基因编码的蛋白质知之甚少。 我们开发了针对p70和hsp70的抗血清来研究 这些蛋白在雄性生殖细胞中的表达和作用。抗人免疫球蛋白抗体 用p70从小鼠精母细胞cDNA文库中筛选克隆。 其中一个克隆与发育中的睾丸特异体杂交 调节3.0kb的mRNA，其表达不受热的影响。数列 分析表明，该克隆与文献报道的同源性较高。 HSP70家族的基因。然而，看起来生精细胞确实 不产生热应激反应的HSP70mRNA或蛋白质。这 提示P70基因的表达与抑制hsp70有关 基因在生精细胞中。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 而P70可能在生殖细胞分化中发挥作用，而不是保护 抵御环境压力的影响。尽管P70的表达可能 有利于精子发生的过程，这一过程也可能 使雄性生殖细胞特别容易受到环境压力的影响。 P70基因的基因组克隆已经分离出来，转基因小鼠将 用来检测生精细胞对 这种基因的表达。

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.