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To investigate the role of SOX9 gene in amphibian gonadogenesis, we analyzed its expression during male and female gonadogenesis in Xenopus tropicalis. The results showed that in both sexes SOX9 mRNA and protein were first detectable after metamorphosis when the gonads were well differentiated and remained present until the adult stage. In the testis, SOX9 expression was restricted to the nucleus of Sertoli-like cells, similarly to what has been observed in other vertebrates suggesting a conserved role in vertebrate testicular differentiation. In the ovary, in sharp contrast with what has been observed in all vertebrates examined so far, the SOX9 protein was localized in the cytoplasm of previtellogenic oocytes before being translocated into the nucleus of vitellogenic oocytes suggesting an unexpected role during oogenesis. These results suggest that the SOX9 gene may not be a sex-determining gene in X. tropicalis and may play different functions in testicular and ovarian differentiation. Developmental Dynamics 237:2996 -3005, 2008.

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The amphibian Xenopus tropicalis, whose genome has been recently sequenced, has become an important model organism for vertebrate developmental genetics. The development of cytogenetic tools in this new model organism should contribute to an understanding of the organization of the amphibian genome and the mapping of a variety of loci of interest. In this respect, oocyte lampbrush chromosomes are particularly useful for the localization of genomic sequences expressed during oogenesis. We have constructed a working map of X. tropicalis lampbrush chromosomes, which allows the 10 bivalents of the oocyte karyotype to be readily identified by distinctive combinations of specific landmark structures composed of lateral loops, spheres, and granules. We have also established the patterns of RNA Pol III sites over the chromosomes by immunofluorescence using antibodies directed against two Pol III subunits. Specific staining patterns were found for each chromosome, which constitute a supplementary tool for their identification.

A proteomic approach has enabled the identification of an orthologue of the splicing factor hnRNP G in the amphibians Xenopus tropicalis, Ambystoma mexicanum, Notophthalmus viridescens and Pleurodeles walt, which shows a specific RNA-binding affinity similar to that of the human hnRN G protein. Three isoforms of this protein with a differential binding affinity for a specific RNA probe were identified in the P. walt oocyte. In situ hybridization to lampbrush chromosomes of P. waltl revealed the presence of a family of hnRNP G genes, which were mapped on the Z and W chromosomes and one autosome. This indicates that the isoforms identified in this study are possibly encoded by a gene family linked to the evolution of sex chromosomes similarly to the hnRNP G/RBMX gene family in mammals.

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