Genetic Changes in Male Infertility

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We have tried to gain insight into the regulatory mechanisms of germ cell development, and to identify genetic changes responsible for male infertility. The results of this project are ;1. (1) We made cDNA libraries of mouse whole testis, purified pachytene spermatocytes and spermatids. (2) From these libraries, we cloned several protein tyrosine phosphatase genes and kinase genes and identified the cells expressing them by in situ hybridization. (3) By doing subtractive hybridization, we isolated genes expressed in the specific stages of the germ cell differentiation.2. To determine the level of gene expression by using clinical biopsy specimens, we developed a non-radioisotopic quantitative RT-PCR method.3. (1) Although both the c-kit protein kinase gene and the mi gene are indispensable for the development of mast cells, we found only the fomer is indispensable for the development of germ cells. (2) We showed that the mi mutant mice have higher frequency of uterine inversion and have less numbers of pups than the wild types.