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Identification of Candidate Genes Involved in Gonocyte Development

ANS M.M. VAN PELT^*,

From the Departments of ^* Biochemistry, Cell Biology, and Histology, Faculty of Veterinary Medicine; and Cell Biology, University Medical Centre Utrecht, Utrecht University, Utrecht, The Netherlands

Correspondence to: Dr A.M.M. van Pelt, Room HP G02.525, Department of Cell Biology, UMC Utrecht, location AZU, Heidelberglaan 100, 3584 CX Utrecht, The Netherlands.

Several germ cell tumors that occur in adult men likely originate from gonocytes that are impaired in their development. To select candidate genes that are involved in the normal development of the gonocytes, we constructed a complementary DNA (cDNA) library from rat gonocytes as well as from single, paired, and aligned A spermatogonia (A_s, A_pr, and A_al spermatogonia, respectively), their direct descendants. Five hundred gonocyte clones were differentially screened using both libraries. Successive verification by dot blot assays yielded 7 clones that were consistently highly abundant only in the gonocyte cDNA library. Also, in situ hybridization of these clones confirmed their differential expression. They encoded for, respectively, succinate dehydrogenase, ribosomal protein S15a, and the 65-kilodalton scaffolding subunit ( isoform) of protein phosphatase 2A. No clues regarding the nature of the remaining four clones could be found. Hence, using differential screening on both constructed cDNA libraries, we were able to select several genes that are interesting candidates for studying the molecular mechanisms of normal gonocyte development.

     Key words: cDNA library, differential screening, in situ hybridization, A spermatogonia, testis, rat

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