Functions and clinical significance of cold shock proteins in the testis

睾丸冷休克蛋白的功能及临床意义

基本信息

批准号：
11470335
负责人：
FUJITA Jun
金额：
$ 6.59万
依托单位：
KYOTO UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B).
财政年份：
1999
资助国家：
日本
起止时间：
1999 至 2000
项目状态：
已结题

项目摘要

Spermatogenesis is a complex process ; a diploid spermatogonium undergoes mitosis and develops into primary spermatocytes. As a result of two meiotic divisions and spermiogenesis, spermatozoa are produced. These processes are controlled by the germ cell - Sertoli cell interactions, and the tightly regulated expression of many genes. In addition to these, spermatogenesis has a unique feature. Unless the testis temperature is maintained to be lower than the body cavity temperature, germ cells will die. To elucidate the molecular mechanisms of spermatogenesis, we tried to isolate genes the expression of which are induced by the testis temperature (32 degree C) and analyze their involvement in male spermatogenesis. The major findings are as follows ; 1) We cloned the mouse cDNA of the cold shock protein, rbm3, and showed that it is expressed in the Sertoli cells of the testis and that its expression is decreased in the cryptorchid testis. 2) By using the yeast hybrid screening method, we tried to identify the RNAs that interact with the cold shock protein Cirp, but no success. 3) We determined the cold shock response element in the DNA sequence of the mouse cirp gene. By using yeast 1-hybrid screening method, we are isolating the protein that binds to this element. 4) Homozygous mutant mice that are deficient in cirp gene were produced. At present no abnormality in phenotypes is apparent. 5) We found that cirp is consticutively expressed the rat neuronal cells and that the expression decreases after transient forebrain ischemia or exposure to H2O2.6) Three proteins that interact with Cirp were isolated by the yeast 2-hybrid screening method. 7) We identified several genes whose expressions are increased in cultured cells by the cold stress.

精子发生是一个复杂的过程。二倍体的精子发生有丝分裂并发展为原发性精子细胞。由于两种减数分裂分裂和精子发生，产生了精子。这些过程由生殖细胞 - 塞托利细胞相互作用以及许多基因的严格调节表达控制。除此之外，精子发生具有独特的特征。除非将睾丸温度保持在低于体腔温度，否则生殖细胞将死亡。为了阐明精子发生的分子机制，我们试图隔离基因，其表达是由睾丸温度诱导的（32度C），并分析了它们参与男性精子发生。主要发现如下； 1）我们将冷休克蛋白RBM3的小鼠cDNA克隆，并表明它在睾丸的Sertoli细胞中表达，并且其表达在Cryptorchid testis中降低。 2）使用酵母杂交筛选方法，我们试图识别与冷休克蛋白CIRP相互作用但没有成功的RNA。 3）我们确定了小鼠CIRP基因的DNA序列中的冷休克响应元件。通过使用酵母1-杂交筛选方法，我们正在分离与该元素结合的蛋白质。 4）产生了CIRP基因不足的纯合突变小鼠。目前，在表型中尚无异常。 5）我们发现CIRP是由组成症表达的大鼠神经元细胞，并且在短暂的前脑缺血或暴露于H2O2.6）三种与CIRP相互作用的蛋白质后的表达降低。 7）我们确定了几个基因，它们的表达在培养的细胞中通过冷应激增加了。