FUNCTION OF THE ter MUTATION AND MOLECULAR CHARACTERISTCS OF A NOVEL PRIMORDIAL GERM CELL GROWTH FACTOR (TERF) IN MICE

小鼠中新型原始生殖细胞生长因子（TERF）的术语突变的功能和分子特征

• 批准号: 09680828
• 负责人: NOGUCHI Motoko
• 金额: $ 2.37万
• 依托单位: SHIZUOKA UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C).
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1998
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09680828/
• 关键词: ter (teratoma) gene; Mouse embryo; Primordial germ cells; Germ-cell growth factor; Germ cell deficiency; Conditioned medium; Gonadal somatic cells; Apoptosis; 細胞増殖因子; TERF; ter遺伝子; コンデイシヨンドメディウム; 生殖細胞欠損; 精巣体細胞; 遺伝子クローニング; コンデイションメデイウム; 生殖細

Congenic mice bearing the ter (teratoma, Chr. 18) mutation that induces the deficiency of primordial germ cells (PGCs) in ter/ter males and females can serve as an animal model of congenital sterility. PGC deficiency occurs in the reconstituted fetal testes containing +/+ PGCs and ter/ter fetal testicular somatic cells. The number of +/+ PGCs that were co-cultured with own somatic cells in the conditioned media (CM) obtained from +/+ and +/ter fetal testicular somatic cells was increased but it was decreased in ter/ter CM. These data suggested a possibility of a novel ter-related growth factor, TERF. In this project the function of the ter gene and molecular characteristics of TERF were analyzed using PGC culture systems and the ter congenic mice. Results obtained were showed below.1. Both +/+ and ter/ter PGCs survived and proliferated on +/+ fetal Sertoli cells co-cultured, but those were degenerated by TUNEL-positive apoptosis on the ter/ter ones, whereas their DNA synthesis occurred … More normally. 2. Administration of +/+ CM from fetal ovarian and testicular somatic cells inhibited apoptosis in +/+ PGCs, but ter/ter CM did not. Both CM supported DNA synthesis of PGCs. 3. Administration of +/+ CM can not inhibit apoptosis in PGCs co-cultured with the ter/ter somatic cells. 4. +/+CM contained heat labile and freeze-resistant protein like substance (m.w. > 30,000) supporting survival of PGCs but not somatic cells. ter/ter CM did not. 5. Several PGC growth factors and their receptors were expressed in ter/terfetal and adult testes as well as in +/+ ones. 6. Thus, it is concluded that a novel ter-related PGC growth factor (designated as TER Factor, TERF) with soluble and membrane-bounded types supports PGC survival by inhibiting apoptosis in PGCs through develdpmental stages and that it is produced by ovarian and testicular somatic cells. It is also concluded that ter/ter somatic cells produce a default type of TERF, resulting in apoptotic death in ter/terPGCs with normal survivability and proliferation ability and sterility of ter/termales and females. 7. We are cloning of the genes in the ter locus now. Less

携带ter（畸胎瘤，Chr. 18）突变的同源小鼠可作为先天性不育的动物模型，该突变诱导ter/ter雄性和雌性中的原始生殖细胞（PGCs）缺乏。PGC缺陷发生在含有+/+ PGC和ter/ter胎儿睾丸体细胞的重建胎儿睾丸中。在+/+和+/ter胎睾丸体细胞的条件培养液中，与自身体细胞共培养的+/+ PGCs数量增加，而在ter/ter条件培养液中，与自身体细胞共培养的+/+ PGCs数量减少。这些数据表明一种新的ter-related生长因子，TERF的可能性。本课题利用PGC培养系统和ter同源小鼠研究了ter基因的功能和TERF的分子特征。所获得的结果如下所示。+/+和ter/ter PGCs在+/+胎儿支持细胞上均能存活并增殖，但在ter/ter细胞上，它们因TUNEL阳性凋亡而退化，而它们的DNA合成则发生在+/+胎儿支持细胞上。 ...更多信息 正常情况下2.给予来自胎儿卵巢和睾丸体细胞的+/+ CM抑制+/+ PGCs的凋亡，但ter/ter CM没有。两种CM均支持PGCs的DNA合成。3. +/+ CM不能抑制与ter/ter体细胞共培养的PGCs的凋亡。4. +/+CM含有热不稳定和抗冻蛋白样物质（m.w. > 30，000）支持PGCs而不是体细胞的存活。ter/ter CM没有。5.几种PGC生长因子及其受体在胎儿和成人睾丸以及+/+睾丸中表达。6.因此，可以得出结论，一种新的ter-related PGC生长因子（命名为TER因子，TERF）具有可溶性和膜结合型支持PGC的生存，通过抑制PGC在发育阶段的凋亡，它是由卵巢和睾丸体细胞产生的。它还得出结论，ter/ter体细胞产生一个默认类型的TERF，导致在ter/terPGCs的凋亡死亡与正常的生存能力和增殖能力和不育的ter/termale和女性。7.我们现在正在克隆ter基因座上的基因。少

项目成果

S.Takabayashi, T.Tokumoto, M.Noguchi, et al.: "Novel growth factor supporting survival of murine primordial germ cells : Evidence from conditioned medium of ter fetal gonadal somatic cells"Molecular Reproduction and Development. 60(3). 384-396 (2001)

S.Takabayashi、T.Tokumoto、M.Noguchi 等人：“支持小鼠原始生殖细胞存活的新型生长因子：来自胎儿性腺体细胞条件培养基的证据”分子生殖和发育。

野口基子: "生殖細胞発生にかかわる突然変異とテラトーマ形成「生殖細胞の発生と性分化」(編著:岡田益吉・長濱嘉孝・中辻憲夫" 共立出版(印刷中), (1998)

野口元子：“与生殖细胞发育和畸胎瘤形成相关的突变”生殖细胞的发育和性分化”（编辑：冈田增吉、长滨义孝、中辻纪夫”共立出版（出版中），（1998）

S.Takabayashi, M.Nozaki, K.Ishikawa, M.Noguchi: "The ter/ter gonadal somatic cells cause apoptosis inter/ter primordial germ cells(PGCs) with normal survivability and proliferation ability in the mouse : Evidence from PGC-somatic cell "exchange-co-culture

S.Takabayashi、M.Nozaki、K.Ishikawa、M.Noguchi：“在小鼠中，三性腺体细胞导致具有正常生存能力和增殖能力的原始生殖细胞 (PGC) 之间的凋亡：来自 PGC 体细胞的证据

野口基子: "生殖細胞発生にかかわる突然変異とテラトーマ形成 「生殖細胞の発生と性分化」 岡田益吉・長濱嘉孝・中辻憲夫 編"共立出版. 9 (1998)

Motoko Noguchi：“与生殖细胞发育和畸胎瘤形成相关的突变。生殖细胞的发育和性分化”，由 Masukichi Okada、Yoshitaka Nagahama 和 Norio Nakatsuji 编辑，Kyoritsu Shuppan 9 (1998)。

Takabayashi, S., Nozaki, M., Ishikawa, K. and Noguchi, M.: "The ter/ter gonadal somatic cells cause apoptosis in ter/ter primordial cells (PGCs) with normal survivability and proliferation ability in the mouse : Evidence from PGC-somatic cell "exchange-co

Takabayashi, S.、Nozaki, M.、Ishikawa, K. 和 Noguchi, M.：“ter/ter 性腺体细胞导致小鼠中具有正常生存能力和增殖能力的 ter/ter 原始细胞 (PGC) 凋亡：证据