Analysis ofSPATA19gene expression during male germ cells development, lessons from in vivo and in vitro study

Abstract

Spermatogenesis-associated-19 (SPATA19) is a novel spermatogenesis related gene that has important biological functions in reproduction. However, the expression aspects of SPATA19 gene are not yet well understood. We analysed SPATA19 gene expression during mouse testis development in vivo (5, 15 and 25 days old mouse testis), in parallel with mouse embryonic stem cells (ESCs) differentiation into male germ-like cells in vitro (5, 11, 19 and 27 days after differentiation), as well as Sertoli cell, mouse embryonic fibroblasts (MEF), and NIH3T3 cancerous cell line. We cultured transgenic mouse ESCs line C57BL/6J expressing Stra8-EGFP in ESC's medium and induced their differentiation by retinoic acid (RA) treatment, and meiotic cells were then sorted by FACS. RT-PCR and real-time PCR were used for analysis of SPATA19 mRNA expression. The expression of SPATA19 mRNA is increased during mouse testis development in vivo and it is highest in 25-days-old mouse testis. SPATA19 is also expressed most highly in mouse ESC-derived germ-like cells after 27 days of RA induction and NIH3T3 cell line but no expression was found in mouse ESC and Sertoli cell line. These findings demonstrate that the expression of SPATA19 increases during male germ cells development. We also suggest an additional possible role for SPATA19 in male germ cell differentiation and tumourigenic process.