| 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 – 2022-03-31
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| キーワード | Spermatogenesis / Mafb / Transcriptions 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 possible treat of infertile patients who are not responding to vitamin A treatment 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 (KO) 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 FY2020, we performed histological analyses of testis from the each single KO mice, and found that MafB and c-Maf expression pattern were changed in both KO mice, indicating that there exist an interaction between the gene expressions. In addition, NGS analyses of testis of MafB conditional KO mice identified several direct target genes involving Vitamin A and MafB signaling.
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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 |
FY2021 is the last 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 |
緊急事態宣言の発出により、共同研究先において外部からの受け入れが出来ずにそこでの実験が実施できなかったため。2021年度中に実施予定である。
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