Identification of novel regulatory genes in development of the avian reproductive tracts.

The chicken reproductive system is unique in maintaining its functions including production of eggs or sperm, fertilization of the egg by sperm maintained in sperm nests, production of hormones regulating its growth, development and function, and reproduction. Development of the reproductive organs is a highly regulated process that results in differentiation and proliferation of germ cells in response to predominant regulatory factors such as hormones and transcription factors. However, only a few genes are known to determine morphogenesis of the chicken reproductive tract and their mechanisms are unknown. Therefore, in the present study, we investigated the expression patterns of four genes including SNCA, TOM1L1, TTR and ZEB1 in the gonads at embryonic days 14 and 18, and in immature (12-week-old) and mature (50-week-old) chickens, as well as the reproductive tract including ovary, oviduct and testes of the respective sexes by qRT-PCR, in situ hybridization and immunofluorescence analyses. The expression of SNCA, TOM1L1 and ZEB1 genes was higher in immature and mature female reproductive tracts than expression of TTR. In addition, different temporal and spatial patterns of expression of the four genes were observed during maturation of testis in chickens. Specifically, SNCA, TOM1L1 and TTR were highly expressed in testes of 12-week-old chickens. Moreover, several chicken specific microRNAs (miRs) were demonstrated to affect expression of target gene mRNAs by directly binding to the 3'-UTR of their target genes through actions at the post-transcriptional level as follows: miR-153 and miR-1643 for SNCA; miR-1680* for TTR; and miR-200b and miR-1786 for ZEB1. These results suggest that four-selected genes play an important role in development of the male and female reproductive tract in chickens and expression of most candidate genes is regulated at the post-transcriptional level through specific microRNAs.