Transgenic Analysis of Gene Governing Reproduction

控制生殖基因的转基因分析

• 批准号: 10044193
• 负责人: NISHIMORI Katsuhiko
• 金额: $ 3.52万
• 依托单位: Tohoku University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B).
• 财政年份: 1998
• 资助国家: 日本
• 起止时间: 1998 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-10044193/
• 关键词: Oxytocin; Oxytocin receptor; GDF-9; DAX-1; Blue mouse; Reproductive organs; Behavioral analysis; MTA-1; FSH-R; OTR; アクチビンβC; アクチビンβE; ノックアウト; C-Kitリガンド

This grant has been a great help for us to set up and carry out those projects listed below. Already, some results has been published, and the other results are close to be published.1) Research on development of mice ovary.1-1. Analysis of anomaly in development of follicles and abnormal proliferation of follicular granulosa cell in gdf-9 -/- mice ovarian development :To further analyze the defect in ovarian follicular development and abnormal proliferation of ovarian granulosa cells of gdf-9-/-mice, we studied the expression pattern of various gene suspected to be directed by gdf-9 and have more direct effect giving rise to the lesion in ovarian follicular development. We showed that the level of the expression of c-kit ligand and hyaluronic acid synthetase might be suppressed in those mutant mice.1-2. Generation of transgenic mice harboring DAX-1-cre :We generated the transgenic mice harboring DAX-1-cre which can facilitate stroma cell specific excision of the target genes such as A … More LK3 , by cre/LoxP mediated recombination in ovaries and testes in mice.2) Analysis of its vivo function of otr and ot gene system by generating gene knockout mice.2-1. Generation of otr gene conditional deficient (knockout) mice :We generated mice whose otr gene was substituted with manipulated otr gene, surrounded by two LoxP sequences (floxed otr).Progeny mice born from ^<flx>otr/^<flx>otr female mice died within 36 hr after birth. This phenotype was suspected to be caused by decreasing level in expression of otr gene in the mutant mice. The quantitation of the level of otr gene expression in these mutant mice were now on going.2-2. Generation of otr gene (conventional knockout) mice :By crossing ^<flx>otr/wt mouse with CAG-cre mice known as general deleter, we obtained conventional otr gene knockout mice. On the contrary to the first expectation, male and female otr-/- mice were so far viable, looked healthy and showed no obvious defect in their appearance. However, viability of progeny mice from the otr heterozygous mice were tend to be lower than those from WT parents.2-3. The system of ot/otr in vivo has remarkable effect on neuronal activity in brain including behavior, learning and memory. After obtaining otr-/- mice at January 2001, we began collaborative research with Dr. Thomas Insel, the director of Center of Behavior and Neuroscience, Emory University, GA, USA.The major purpose of this collaboration is to analyze the abnormal behavior in otr-/- mice, and to use these mutant mice as a material to analyze the function of otr gene ectopically expressed by virus vectors in mice brain.3) Functional Analysis of MTA-1 gene.3-1 MTA-1 gene is showing strong correlation with metastasis and malignancy of cancer cell derived from reproductive organs such as ovary and mammary gland. We reported the effect of MTA-1 gene expression on cell proliferation, on the cell's metastatic ability and on the transcriptional activity of GATA enhancer element. Less

这笔赠款对我们建立和实施下列项目有很大的帮助。已经有一些结果已经发表，其他结果也即将发表。1）小鼠卵巢发育的研究。1 -1。gdf-9 -/-小鼠卵巢发育中卵泡发育异常和卵泡颗粒细胞异常增殖的分析：为了进一步分析gdf-9-/-小鼠卵巢卵泡发育缺陷和卵巢颗粒细胞异常增殖，我们研究了各种疑似由gdf-9指导的、对引起卵巢卵泡发育病变有更直接影响的基因的表达模式。我们发现，c-kit配体和透明质酸合成酶的表达水平可能在这些突变小鼠中受到抑制。1 -2.携带DAX-1-cre的转基因小鼠的产生：我们产生了携带DAX-1-cre的转基因小鼠，其可以促进基质细胞特异性切除靶基因，如A ...更多信息 LK 3，通过cre/LoxP介导的重组在小鼠的卵巢和睾丸中。2）通过产生基因敲除小鼠分析其对otr和ot基因系统的体内功能。otr基因条件性缺陷（敲除）小鼠的产生：我们产生了用被两个LoxP序列（floxed otr）包围的经操纵的otr基因取代otr基因的小鼠<flx><flx>。这种表型可能是由于突变小鼠otr基因表达水平降低所致。这些突变小鼠中otr基因表达水平的定量研究正在进行中。otr基因（常规敲除）小鼠的产生：通过将<flx>otr/wt小鼠与称为通用删除者的CAG-cre小鼠杂交，我们获得了常规otr基因敲除小鼠。与第一种预期相反，雄性和雌性otr-/-小鼠到目前为止都是可行的，看起来很健康，外观上没有明显的缺陷。然而，来自otr杂合小鼠的子代小鼠的存活率倾向于低于来自WT亲本的子代小鼠。在体OT/OTR系统对脑内神经元活动包括行为、学习和记忆有显著影响。2001年1月获得otr-/-小鼠后，我们开始与美国佐治亚州埃默里大学行为与神经科学中心主任托马斯·英塞尔博士合作研究，主要目的是分析otr-/-小鼠的异常行为，并以这些突变小鼠为材料，分析病毒载体异位表达的otr基因在小鼠脑中的功能。3 -1 MTA-1基因显示出与来自生殖器官如卵巢和乳腺的癌细胞的转移和恶性化的强相关性。我们报道了MTA-1基因表达对细胞增殖、转移能力和加塔增强子转录活性的影响。少

项目成果

Lau,A.et al.: "Activin bC and activin bE genes are not essential for mouse liver growth, differentiation, and regeneration."Mol.Cell Biol.. 20. 6127-6137 (2000)

Lau,A.等人：“激活素 bC 和激活素 bE 基因对于小鼠肝脏生长、分化和再生不是必需的。”Mol.Cell Biol.. 20. 6127-6137 (2000)

Nomura,O. et al.: "Expression of five steroidogenic genes including aromatase gene at early developmental stage of chicken male and female embryos."J.Steroid Biochem.Mol.Biol.. 71. 103 (1999)

野村，O.

Nawa,A. et al.: "Tumor metastasis-associated human MTA1 gene : its sequence analysis and association with cancer cell proliferation."Cell Biochem.. 79. 202 (2000)

名和，A.

Nomura,O. et al.: "Expression of five steroidogenic genes including aromatase gene at earlydevelopmental stage of chicken male and female embryos."J.Steroid Biochem.Mol.Biol.. 71. 103-109 (1999)

野村，O.

Lau, A.L., Kumar, T.R., Nishimori, K., Bonadio, J., and Matzuk, M.M.: "Activin bC and activin bE genes are not essential for mouse liver growth, differentiation, and regeneration."Mol.Cell Biol. 20. 6127-6137 (2000)

Lau, A.L.、Kumar, T.R.、Nishimori, K.、Bonadio, J. 和 Matzuk, M.M.：“激活素 bC 和激活素 bE 基因对于小鼠肝脏生长、分化和再生不是必需的。”Mol.Cell Biol。