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• W1197230070 abstract "During mammalian pre-implantation development pluripotent cells are established within the inner cellmass (ICM). These pluripotent cells contribute to all the somatic cell lineages and establish the germline, the only cell lineage to be transmitted to the next generation. The molecular mechanisms leadingto germ cell specification have not been elucidated, but some genes that are critical for pluripotent cellsand during pre-implantation development are subsequently restricted to the germ lineage. Among thesegenes Sox2 has been shown to be specifically expressed in the ICM, in neural stem cells and inprimordial germ cells (PGCs). Sox2 deletion is lethal very early during embryogenesis, and nullblastocysts do not develop correctly at implantation.To understand the timing of Sox2 expression in germ cells, we first performed β-galactosidasehistochemistry on whole-mount embryos, including gonads, at various developmental stages by usingSox2-βgeo knock-in mice. By Northern blot analysis we found Sox2 expression in postnatalspermatogonia and testes, from 1 dpp up to the puberal age. Since it has been shown, by RNA wholemountin situ hybridization, that Sox2 is expressed in PGCs, we performed an immunohistochemistry(IHC) on testis sections. Interestingly, we found Sox2 expression in 13 dpc pro-spermatogonia as wellas in spermatogonial stem cells. This suggests that, Sox2 expression in male germline is associated tothe stem cell compartment. In female germ cells, we found positivity in growing oocytes and in 1 dpcfertilized egg by immunofluorescence (IF). We also found that Sox2 is expressed in the human embryoin the developing neural tube, neural crest cells and in scattered cells, between the dorsal mesentery andthe mesonephroi, which were positive for Oct4 and suggestive of PGCs. Sox2 is also expressed in fetallung and after, in lung and thyroid cancers of human adult.To specifically target Sox2 deletion in the germ line, Sox2-lox mice were first cross bred to Sox2-βgeo/TNAP-Cre mice, to generate Sox2lox/Sox2βgeo/TNAP-Cre animals. Sox2 excision in the gonadswas verified by PCR, western blot and IHC. Deleted female and male 12 dpc embryos were thensacrificed and gonads analysed. By PCR genotyping the gonads deletion was confirmed. However,immunostaining of the deleted gonads showed Sox2 expression, suggesting probably, that Cremediatedexcision of Sox2 was only partial. When a deleted male was mated to wt female we observednormal litter size, but without the Sox2null allele, suggesting a potential role for Sox2 in male gametesmaturation. When a deleted adult female was mated to a wt male, a strong reduction of the litter sizewas observed. Analysis of 2 dpc embryos, obtained from this intercross, outgrown in vitro for 2.5 daysshowed that they did not develop beyond 2/4-cell stage and were negative, by IF, to Sox2 and Oct4staining. This finding suggests that Sox2 could also be important, in female gametes, for the earlyembryonic development." @default.
• W1197230070 created "2016-06-24" @default.
• W1197230070 creator A5027644299 @default.
• W1197230070 date "2008-05-28" @default.
• W1197230070 modified "2023-09-23" @default.
• W1197230070 title "Sox2 conditional deletion in germ cells: a potential role in gametogenesis" @default.
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