Conserved role of Ovo in germline development in mouse and Drosophila

Hayashi, Makoto; Shinozuka, Yuko; Shigenobu, Shuji; Sato, Mitsuru; Sugimoto, Michihiko; Ito, Seiji; Abe, Kuniya; Kobayashi, Satoru

doi:10.1038/srep40056

Cited by 29 publications

(40 citation statements)

References 46 publications

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“…; Hayashi et al . ). The embryos expressing Sxl‐EGFP in PGCs at stages 10–15 were collected for RIP‐seq analysis with anti‐EGFP antibody.…”

Section: Resultsmentioning

confidence: 97%

“…; Hayashi et al . ). Among the seven aforementioned genes, four were required for germline development.…”

Section: Resultsmentioning

confidence: 97%

“…Whole EGFP-vasa embryos were also collected at 11 different stages. Total RNA extraction, RNA amplification, and microarray analysis were carried out as described (Hayashi et al 2017). The resultant data from PGCs and whole embryos were deposited in GEO under Accession No.…”

Section: Microarray Analysismentioning

confidence: 99%

“…To express Sxl-EGFP in PGCs, we used nos-Gal4-VP16 (nos-Gal4), which produces Gal4-VP16 under the control of the germline-specific nos promoter (Van Doren et al 1998). In embryos produced by females carrying the nos-Gal4 gene (maternal nos-Gal4), maternally supplied nos-Gal4 RNA is partitioned into PGCs, and its protein product activates UAS-dependent gene expression from stage 9 until at least the end of embryogenesis (Van Doren et al 1998;Hayashi et al 2017). The embryos expressing Sxl-EGFP in PGCs at stages 10-15 were collected for RIP-seq analysis with anti-EGFP antibody.…”

Section: Isolation Of Rnas Immunoprecipitated With Sxl Protein In Pgcsmentioning

confidence: 99%

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Transcripts immunoprecipitated with Sxl protein in primordial germ cells of Drosophila embryos

et al. 2017

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In Drosophila, Sex lethal (Sxl), an RNA binding protein, is required for induction of female sexual identity in both somatic and germline cells. Although the Sxl-dependent feminizing pathway in the soma was previously elucidated, the downstream targets for Sxl in the germline remained elusive. To identify these target genes, we selected transcripts associated with Sxl in primordial germ cells (PGCs) of embryos using RNA immunoprecipitation coupled to sequencing (RIP-seq) analysis. A total of 308 transcripts encoded by 282 genes were obtained. Seven of these genes, expressed at higher levels in PGCs as determined by microarray and in situ hybridization analyses, were subjected to RNAi-mediated functional analyses. Knockdown of Neos, Kap-alpha3, and CG32075 throughout germline development caused gonadal dysgenesis in a sex-dependent manner, and Su(var)2-10 knockdown caused gonadal dysgenesis in both sexes. Moreover, as with knockdown of Sxl, knockdown of Su(var)2-10 in PGCs gave rise to a tumorous phenotype of germline cells in ovaries. Because this phenotype indicates loss of female identity of germline cells, we consider Su(var)2-10 to be a strong candidate target of Sxl in PGCs. Our results represent a first step toward elucidating the Sxl-dependent feminizing pathway in the germline.

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“…; Hayashi et al . ). The embryos expressing Sxl‐EGFP in PGCs at stages 10–15 were collected for RIP‐seq analysis with anti‐EGFP antibody.…”

Section: Resultsmentioning

confidence: 97%

“…; Hayashi et al . ). Among the seven aforementioned genes, four were required for germline development.…”

Section: Resultsmentioning

confidence: 97%

Section: Microarray Analysismentioning

confidence: 99%

Section: Isolation Of Rnas Immunoprecipitated With Sxl Protein In Pgcsmentioning

confidence: 99%

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Transcripts immunoprecipitated with Sxl protein in primordial germ cells of Drosophila embryos

et al. 2017

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“…These transgenes were transcribed in PGCs using the nos-Gal4-VP16 (nos-Gal4) transgene, which expresses the transcriptional activator, Gal4-VP16, under the control of the germline-specific nos promoter (Van Doren et al 1998). In PGCs, Gal4-VP16 produced from the maternal transcript (maternal nos-Gal4) activated UAS-dependent gene expression from stage 11 until at least the end of embryogenesis (Hayashi et al 2017). In PGCs, Gal4-VP16 produced from the maternal transcript (maternal nos-Gal4) activated UAS-dependent gene expression from stage 11 until at least the end of embryogenesis (Hayashi et al 2017).…”

Section: Cg32425 3 0 Utr Mediated Nos-dependent Upregulation Of Rna Amentioning

confidence: 99%

Maternal Nanos‐Dependent RNA Stabilization in the Primordial Germ Cells of Drosophila Embryos

2017

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Nanos (Nos) is an evolutionary conserved protein expressed in the germline of various animal species. In Drosophila, maternal Nos protein is essential for germline development. In the germline progenitors, or the primordial germ cells (PGCs), Nos binds to the 3' UTR of target mRNAs to repress their translation. In contrast to this prevailing role of Nos, here we report that the 3' UTR of CG32425 mRNA mediates Nos-dependent RNA stabilization in PGCs. We found that the level of mRNA expressed from a reporter gene fused to the CG32425 3' UTR was significantly reduced in PGCs lacking maternal Nos (nos PGCs) as compared with normal PGCs. By deleting the CG32425 3' UTR, we identified the region required for mRNA stabilization, which includes Nos-binding sites. In normal embryos, CG32425 mRNA was maternally supplied into PGCs and remained in this cell type during embryogenesis. However, as expected from our reporter assay, the levels of CG32425 mRNA and its protein product expressed in nos PGCs were lower than in normal PGCs. Thus, we propose that Nos protein has dual functions in translational repression and stabilization of specific RNAs to ensure proper germline development.

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Disruption of the ovarian serine protease (Osp) gene causes female sterility in Bombyx mori and Spodoptera litura

Wang

et al. 2019

Pest Management Science

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BACKGROUND Precise regulation of oogenesis is crucial to female reproduction. Seventy percent of pests belong to lepidopteran species, so it would be interesting to explore the highly conserved genes involved in oogenesis that do not affect growth and development in the lepidopteran model, Bombyx mori. This can provide potential target genes for pest control and promote the development of insect sterility technology. RESULTS In lepidopteran species, ovarian serine protease (Osp), which encodes a member of the serine protease family, is essential for oogenesis. In this study, we used transgenic CRISPR/Cas9 technology to obtain Osp mutants in the model lepidopteran insect Bombyx mori and in the lepidopteran agricultural pest Spodoptera litura. Sequence analysis of mutants revealed an array of deletions in Osp loci in both species. We found that the deletion of Osp resulted in female sterility, whereas male fertility was not affected. Although B. mori and S. litura mutant females mated normally, they laid fewer eggs than wild‐type females and eggs did not hatch. CONCLUSION Osp is crucial for female reproductive success in two species of Lepidoptera. As the Osp gene is highly conserved in insect species, this gene is a potential molecular target for genetic‐based pest management. © 2019 Society of Chemical Industry

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Conserved role of Ovo in germline development in mouse and Drosophila

Cited by 29 publications

References 46 publications

Transcripts immunoprecipitated with Sxl protein in primordial germ cells of Drosophila embryos

Transcripts immunoprecipitated with Sxl protein in primordial germ cells of Drosophila embryos

Maternal Nanos‐Dependent RNA Stabilization in the Primordial Germ Cells of Drosophila Embryos

Disruption of the ovarian serine protease (Osp) gene causes female sterility in Bombyx mori and Spodoptera litura

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