Genome‐wide identification of Sox8‐, and Sox9‐dependent genes during early post‐natal testis development in the mouse

Abstract

Genome-wide identification of Sox8-, and Sox9-dependent genes during early post-natal testis development in the mouse F. Chalmel,*,a A. Lardenois,*,a I. Georg, †, ‡,k F. Barrionuevo, †,** P. Demougin, § B. J egou,*, ¶ G. Scherer † and M. Primig* *Inserm, U1085-Irset, University of Rennes 1, Rennes, France, †Institute of Human Genetics, University of Freiburg, Freiburg, Germany, ‡Faculty for Biology, University of Freiburg, Freiburg, Germany, §Biozentrum, University of Basel, Basel, Switzerland, ¶EHESP School of Public Health (Rennes, Paris), Rennes Cedex, France, k Present address: Area of Human DNA Variability, Centro de Genomica e Investigacio Oncologica (GENYO), Pfizer-Universidad de Granada-Junta de Andalucia, Granada, Spain, and **Departamento de Genetica e Instituto de Biotecnologia, Universidad de Granada, Armilla, Spain SUMMARYThe SOX8 and SOX9 transcription factors are involved in, among others, sex differentiation, male gonad development and adult maintenance of spermatogenesis. Sox8 À/À mice lacking Sox9 in Sertoli cells fail to form testis cords and cannot establish spermatogenesis. Although genetic and histological data show an important role for these transcription factors in regulating spermatogenesis, it is not clear which genes depend upon them at a genome-wide level. To identify transcripts that respond to the absence of Sox8 in all cells and Sox9 in Sertoli cells we measured mRNA concentrations in testicular samples from mice at 0, 6 and 18 days post-partum. In total, 621 and 629 transcripts were found at decreased or increased levels, respectively, at different time points in the mutant as compared to the control samples. These mRNAs were categorized as preferentially expressed in Sertoli cells or germ cells using data obtained with male and female gonad samples and enriched testicular cell populations. Five candidate genes were validated at the protein level. Furthermore, we identified putative direct SOX8 and SOX9 target genes by integrating predicted SOX-binding sites present in potential regulatory regions upstream of the transcription start site. Finally, we used protein network data to gain insight into the effects on regulatory interactions that occur when Sox8 and Sox9 are absent in developing Sertoli cells. The integration of testicular samples with enriched Sertoli cells, germ cells and female gonads enabled us to broadly distinguish transcripts directly affected in Sertoli cells from others that respond to secondary events in testicular cell types. Thus, combined RNA profiling signals, motif predictions and network data identified putative SOX8/SOX9 target genes in Sertoli cells and yielded insight into regulatory interactions that depend upon these transcription factors. In addition, our results will facilitate the interpretation of genome-wide in vivo SOX8 and SOX9 DNA binding data.

Expression of SOX3 and SOX9 Genes in Gonads of Blue Gourami

“…The results of this study are in agreement with a previous finding that SOX9 is present in both sexes, but in one of the first molecular events in the male pathway is upregulated in Sertoli cell precursors (which are essential for testis differentiation) by the primary sex-determining trigger interacting with an evolutionarily conserved sequence within SOX9 [29]. In mammals, the SOX8 and SOX9 transcription factors are involved, among other things, in sex differentiation, male gonad development and adult maintenance of spermatogenesis [30].…”

Gonadal Identity in the Absence of Pro-Testis Factor SOX9 and Pro-Ovary Factor Beta-Catenin in Mice1

“…While Sox8 inactivation does not affect testis determination [47], analyses of Sox8/Sox9 double mutants showed that SOX8 is able to compensate for the loss of Sox9 after sex determination [9,30,48]. Moreover, SOX8 and SOX9 activate common target genes such as Amh and Cbln4 [32,48]. Therefore, we propose that SRY and its target gene SOX8 partially compensate for the absence of SOX9 in XY Ctnnb1/Sox9 DKO supporting cells and promote Sertoli cell differentiation (Fig.…”

“…In our XY Ctnnb1/Sox9 DKO model, in addition to Sry, Sox8 is the only Sox gene expressed during the initial events of masculinization. While Sox8 inactivation does not affect testis determination [47], analyses of Sox8/Sox9 double mutants showed that SOX8 is able to compensate for the loss of Sox9 after sex determination [9,30,48]. Moreover, SOX8 and SOX9 activate common target genes such as Amh and Cbln4 [32,48].…”

RNAi silencing of the SoxE gene suppresses cell proliferation in silkworm BmN4 cells

“…The direct transcriptional targets of Sox10 include genes encoding proteolipid protein, extracellular superoxide dismutase, and pleiotrophin in rat Schwannoma cells [10]. Moreover, genome-wide analysis has revealed hundreds of genes that are potential binding targets for Sox9 and/or Sox8 in mice and rats [11, 12]. Because of the functional redundancy of the different SoxE proteins in mammals [13], it may be difficult to determine their targets.…”