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Visualization of gonadal development and sex differentiation in medaka using in vivo imaging

使用体内成像观察青鳉性腺发育和性别分化

基本信息

批准号：
18570064
负责人：
SAITO Daisuke
金额：
$ 2.14万
依托单位：
National Institute for Basic Biology
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2006
资助国家：
日本
起止时间：
2006 至 2007
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-18570064/
关键词：
Development and differentiation Gonadogenesis Sex differentiation Medaka Imaging

项目摘要

This project aimed to depict the cellular interaction of cell lineages involved in the gonadal sex differentiation and development in the medakafish by applying recently developed BAC transgenic technology for genes specifically expressed in developing gonads. We successfully generated the BAC transgenic lines for several gonad-specific transcription factors such as sox9b ftz-f1, and pod1, and the medaka sex determination factor dmy; steroidgenesis enzyme p450c17, male-specific somatic marker gene dmrt1, and female-specific marker gene aromatase. For each transgenic line, we examined the temporal and spatial expression patterns of transgene-expressing cells by a whole-mount immunofluorescence method. Using sox9b-EGFP transgenic line, we found that sox9b-expressing cells are common precursors of both testicular and ovarian supporting cells (Nakamura et al, 2008). We also applied the BAC technology for the analysis of mutant line zenzai, which showed developmental germline depletion phen … More otype. Using the transgenic line for candidate gene zenzai and rescue line expressing zenzai-EGFP fusion gene, we examined the developmental dynamics of zenzai-expressing cells. Further, by crossing sox9b-EGFP line or olvas-EGFP line with zenzai mutant line, we found the firm relationship between pattern of germ cell division and mutant phenotype (Saito et al, 2007). Another major finding includes the analysis of germ cell division in adult ovarian surface using the time-lapse imaging of transgene-expressing cells. Double-color transgenic line turned out to be especially useful for such purposes. We found the putative germline stem cells in adult ovarian tissue (Nakamura et al, unpublished). Taken together, we found the several important aspects of cellular interaction during gonadal development in medaka, which showed the potential usefulness of BAC transgenic technology in developmental and reproductive biology. The newly generated transgenic lines will be valuable resources for further research. Less

本项目旨在通过应用新近开发的BAC转基因技术，研究青鳉鱼性腺发育中特异表达基因的细胞间相互作用。我们成功地产生了BAC转基因株系的几个性腺特异性转录因子，如sox 9 b ftz-f1和pod 1，青鳉性别决定因子dmy，类固醇生成酶p450 c17，男性特异性体细胞标记基因dmrt 1，和女性特异性标记基因芳香化酶。对于每一个转基因株系，我们研究了时间和空间的表达模式的转基因表达细胞的整体安装免疫荧光法。使用sox 9 b-EGFP转基因系，我们发现表达sox 9 b的细胞是睾丸和卵巢支持细胞的共同前体（中村等人，2008）。我们还应用BAC技术分析了突变系zenzai，该突变系表现出发育性种系缺失phen， ...更多信息类型。使用候选基因zenzai的转基因株系和表达zenzai-EGFP融合基因的拯救株系，我们检查了表达zenzai的细胞的发育动态。此外，通过将sox 9 b-EGFP系或olvas-EGFP系与zenzai突变系杂交，我们发现了生殖细胞分裂模式和突变表型之间的牢固关系（Saito等，2007）。另一个主要发现包括使用转基因表达细胞的延时成像分析成人卵巢表面的生殖细胞分裂。双色转基因系被证明是特别有用的，为这些目的。我们在成人卵巢组织中发现了假定的生殖系干细胞（中村等，未发表）。综上所述，我们发现了青鳉性腺发育过程中细胞间相互作用的几个重要方面，这表明BAC转基因技术在发育和生殖生物学中的潜在用途。新获得的转基因株系将为进一步的研究提供宝贵的资源。少