| Budget Amount *help |
¥3,340,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥240,000)
Fiscal Year 2007: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
Fiscal Year 2006: ¥2,300,000 (Direct Cost: ¥2,300,000)
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| Research Abstract |
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
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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 We found the putative germline stem cells in adult ovarian tissue (Nakamura et al, unpublished).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
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