| 研究課題/領域番号 |
19K16027
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| 研究機関 | 名古屋市立大学 |
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研究代表者 |
シャウキ ホッサム 名古屋市立大学, 医薬学総合研究院(医学), 助教 (70829738)
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| 研究期間 (年度) |
2019-04-01 – 2021-03-31
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| キーワード | spermatogenesis / Mafb / transcription factor |
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| 研究実績の概要 |
The project aimed to identify the vitamin A downstream targets that lead to induction of spermatogonial stem cell differentiation, which will contribute to treating infertile patients who are not responding to vitamin A treatment itself due to mutated downstream targets. In FY2019, we identified two vitamin A targets; MafB and c-Maf. We then generated the floxed alleles of each gene independently and then lines were mated to obtain double conditional knockout mice, in which Cre recombinase was activated upon tamoxifen treatment. We induced Cre activation at 6 weeks old mice and mice tissues were collected and the deletion of both genes were confirmed. In the second year of this project, we are going to analyze the fertility of the 2 Mo (young) and 8 Mo (old) male mice upon deletions.
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| 現在までの達成度 (区分) |
現在までの達成度 (区分)
2: おおむね順調に進展している
理由
The plan of this research are to (1) generate the MafB/c-Maf single and double knockout mice, (2) analyzing whether Mafb/c-Maf compensate each other to fully induce the first spermatogenetic cycle under the control of retinoic acid using the KO mice, (3) rescue vitamin A deficient mouse phenotype by lentivirus overexpressing of Mafb/c-Maf. The first task was achieved during the first year (FY2019) while the second and third tasks are going to be elucidated during the second year (FY2020). the project was progressing rather smoothly in FY2019 to generate the double cKO. However, we noticed that the DKO developed urinary proteinuria after 2 weeks of Cre activation, a sign of kidney dysfunction, therefore we are also aiming to analyze the kidney abnormality caused by Mafb/c-Maf deletion.
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| 今後の研究の推進方策 |
FY2020 is the second year of the research, and we will promote the research as following (1) analyze the physiology and histology of single and double MafB/c-Maf cKO testis, (2) perform fertility test of the KO males, (3) perform RNA-seq from the DKO Sertoli cells to identify the secreted proteins regulated by Maf transcription factors, (4) rescue vitamin A deficient mouse testis by overexpressing of Maf factors. (5) analyze the kidney abnormality caused by double Mafb/c-Maf deletions.
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| 次年度使用額が生じた理由 |
令和元年度に予定したin vitroの実験が一部令和2年度にずれ込んだため。ただし、全体の実験計画に影響はなく、令和2年度末までに、予定の研究を終えることができる見込みである。
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