Nanos2-associated mRNAs in the postnatal testis
Ontology highlight
ABSTRACT:
ORGANISM(S): Mus musculus
PROVIDER: GSE61807 | GEO | 2015/09/25
SECONDARY ACCESSION(S): PRJNA262171
REPOSITORIES: GEO
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Project description:To identify Nanos2-associated mRNAs, we performed microarray analysis of mRNAs coprecipitated with FLAG-tagged NANOS2 with male gonad from Tg-Nanos2 enhancer-3xFLAG-Nanos2-3'UTR at E14.5.
2013-02-06 | GSE37718 | GEO
Project description:To identify Nanos2-associated mRNAs, we performed microarray analysis of mRNAs coprecipitated with FLAG-tagged NANOS2 with male gonad from Tg-Nanos2 enhancer-3xFLAG-Nanos2-3'UTR at E14.5. Biological duplicates were examined at each sample.
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Project description:To investigate the biochemical function of NANOS2, we performed expression microarray analysis of the embyornic male gonad of Nanos2 hetero, Nanos2 KO, and Nanos2 KO_Tg 3×FLAG-tagged Nanos2-ΔN10, which is truncated form of Nanos2 in the N-terminal region, transgenic mice.
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Project description:Spermatogonial stem cells are foundation of spermatogenesis. In our previous study, we observed a rapid degeneration of spermatogenesis by conditional knockout (cKO) of Nanos2 in adult spermatogonia, but the underlying mechanisms are largely unknown. While, conditional overexpression (cOE) of Nanos2 also inhibited germ cell development and stayed in undifferentiation status. Here we used cultured germline stem cells (GSCs) to study the mechanisms of Nanos2 in spermatogonia. We compared Nanos2 expression in cultured GSCs with in vivo isolated Nanos2 positive cells, and found the level of Nanos2 is significantly lower in GSCs as compared to Nanos2 positive cells. Thus we generated Nano2 cOE GSCs lines. By conditional inducing expression of Nanos2 we compared gene expression profile to control GSCs. These data provide the first evidence of Nanos2 regulating genes in spermatogonial stem cells.
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Project description:In mouse embryos, sex-specific differentiation of primordial germ cells (PGCs) begins with distinct cell cycle regulations: while PGCs in female (XX) gonads enter meiosis, PGCs in male (XY) gonads defer meiosis and arrest cell cycle1,2. The mitotic quiescence in XY gonads requires Nanos2, an evolutionarily conserved RNA-binding protein implicated in RNA degradation3-5; however, underlying mechanisms including the identity of its target mRNAs in vivo remain unknown. Here we show that Nanos2 negatively regulates Dazl (Deleted in azoospermia-like), a germline-specific gene encoding an RNA-binding protein that is implicated in translational control and meiotic initiation6,7. Comprehensive microarray analyses identified Dazl as one of the genes that were up-regulated in Nanos2-deficient XY gonads, but down-regulated in Nanos2-expressing XX gonads. Analysis of a BAC transgenic mouse line, in which the 3’UTR of Dazl can be removed by Flp-Frt recombination system, showed that repression of Dazl is likely achieved by the association of Nanos2 with Dazl’s 3’UTR. Elevated levels of Dazl expression in the transgenic XY PGCs resulted in abnormal progression of cell cycles including meiosis, reminiscent of the phenotype observed in Nanos2-/- XY gonads. This phenotype was suppressed by inhibiting retinoic acid (RA) signaling, suggesting that Nanos2 represses Dazl to prevent RA-induced cell cycle progression in XY PGCs. Furthermore, we show that a part of Nanos2-associated mRNAs, most of which are up-regulated in Nanos2-/- XY gonads, can also associate with Dazl, and that excess Dazl impedes localization of Nanos2 to processing-bodies where RNA degradation takes place, suggesting that repression of Dazl is required for Nanos2 to efficiently degrade its target mRNAs. These data provide the first link between the two RNA-binding proteins that play pivotal roles in sexual differentiation of murine PGCs.
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Project description:In mouse embryos, sex-specific differentiation of primordial germ cells (PGCs) begins with distinct cell cycle regulations: while PGCs in female (XX) gonads enter meiosis, PGCs in male (XY) gonads defer meiosis and arrest cell cycle1,2. The mitotic quiescence in XY gonads requires Nanos2, an evolutionarily conserved RNA-binding protein implicated in RNA degradation3-5; however, underlying mechanisms including the identity of its target mRNAs in vivo remain unknown. Here we show that Nanos2 negatively regulates Dazl (Deleted in azoospermia-like), a germline-specific gene encoding an RNA-binding protein that is implicated in translational control and meiotic initiation6,7. Comprehensive microarray analyses identified Dazl as one of the genes that were up-regulated in Nanos2-deficient XY gonads, but down-regulated in Nanos2-expressing XX gonads. Analysis of a BAC transgenic mouse line, in which the 3’UTR of Dazl can be removed by Flp-Frt recombination system, showed that repression of Dazl is likely achieved by the association of Nanos2 with Dazl’s 3’UTR. Elevated levels of Dazl expression in the transgenic XY PGCs resulted in abnormal progression of cell cycles including meiosis, reminiscent of the phenotype observed in Nanos2-/- XY gonads. This phenotype was suppressed by inhibiting retinoic acid (RA) signaling, suggesting that Nanos2 represses Dazl to prevent RA-induced cell cycle progression in XY PGCs. Furthermore, we show that a part of Nanos2-associated mRNAs, most of which are up-regulated in Nanos2-/- XY gonads, can also associate with Dazl, and that excess Dazl impedes localization of Nanos2 to processing-bodies where RNA degradation takes place, suggesting that repression of Dazl is required for Nanos2 to efficiently degrade its target mRNAs. These data provide the first link between the two RNA-binding proteins that play pivotal roles in sexual differentiation of murine PGCs.
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Project description:To investigate the biochemical function of NANOS2, we performed expression microarray analysis of the embyornic male gonad of Nanos2 hetero, Nanos2 KO, and Nanos2 KO_Tg 3M-CM-^WFLAG-tagged Nanos2-M-NM-^TN10, which is truncated form of Nanos2 in the N-terminal region, transgenic mice. 3M-CM-^WFLAG-tagged Nanos2-M-NM-^TN10 protein function was validated by rescue experiment in the Nanos2-null male gonad of mouse embryo at E14.5. Biological duplicates were examined at each genotype, Nanos2 hetero, Nanos2 KO, and Nanos2 KO_Tg 3M-CM-^WFLAG-tagged Nanos2-M-NM-^TN10 for each experiment.
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Project description:Gene expression of Tg 3×FLAG-Nanos2-deltaN10 mouse male gonad
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