Analysis of Meiosis using Hybrid-male-sterile Animals.

使用杂交雄性不育动物进行减数分裂分析。

• 批准号: 05044128
• 负责人: HOTTA Yasuo
• 金额: $ 3.84万
• 依托单位: Nara Institute of Science and Technology (1994)Nagoya University (1993)
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for international Scientific Research
• 财政年份: 1993
• 资助国家: 日本
• 起止时间: 1993 至 1994
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-05044128/
• 关键词: Meiosis; Male-sterile; testes; Ovaries; RecA; Lim15; DNA polymerase; apoptosis; 精母細胞; シナプトネマ構造; 染色体対合; 核構造; ヤク; シカ

Meiosis is one of the important process for formation of the functionally reproductive cells and the major events are the piring of homologous chromosomes, genetic recombination (crossing-over) and chiasma-formation between them and segregation of chromosomes (disjunction).The failure of any one of these events leads to lack of reproductive cells and sterility. Despite the accumulation of many cytological and cytogenetical studies on meiosis, molecular analyzes of meiotic events are just underway using yeasts, but still scarece in higher organisms. Nevertheless, understanding and control of meiotic events could make uncountable contribution for breeding, fertility and sterility problems and human welfare. We have tried to analyze testicular and ovarian cells from the male-sterile animals, Yak (F1 of Bos taurus X Bos grunniens), Cerus elaphus X Cerus nippon F1 (deer), M.fascicularis X M.assamesis F1 (monkey), cytologically and molecular biologically using our antibodies to recombination … More al enzymes and various probes prepared from the meiotically active genes. Currently we have completed our studies using yaks. The hybrid-male-sterile deers are not yet in our hand, while hybrid-sterile-monkey died and the next production is questionable in Kunming Institute. The followings are our experimental results today :1.The male-sterility is caused by meiotic degradation mostly during pchytene and some at diakinesis and metaphase I.However, the similar meiotic degradation has been observed in oocytes, suggesting the reduction in number of the oocytes. The antibody prepared against RecA protein and Lim15 (lily meiotic prophase I-specific gene) protein using E.coli have positively bound to most of meiotic cells even from sterile testes, while little binding has been observed in somatic cells including the testicular and the ovarian ones.2.Acivities of DNA polymerase beta and alpha isolated from meioic prophase cells of sterile and fertile (normal ox) animal are essentially the same, based on the cellular protein.3.Isolated cDNA clones from sterile and fertile testicular meiotic prophase cells are similar in restriction maps and quantities obtained, suggesting that the normal transcription is occurring in meiotic prophase cells from the sterile males.All of these can gives us a clue to conclude the following but it is preliminary and further carefull studies should be carried out.1.So-called male-sterility is caused by the meiotic break down lead to reduction of sperm. This occurs also in female. Since female fertility is obtainable with one egg cell, the female is fertile despite the reduction in the number of oocytes.2.The molecular events so far examined indicated that there is no detectable difference between the fertiles and the steriles until either prophase or metaphase. The meiotic degradation in these hybrids may be due to apoptosis as speculated in mutant mouse, but this requirs more analysis. Less

减数分裂是形成具有生殖功能的细胞的重要过程之一，其主要事件是同源染色体的纯化、染色体间的遗传重组（交换）和交叉形成以及染色体分离（分离），任何一个事件的失败都会导致生殖细胞的缺乏和不育。尽管对减数分裂的细胞学和细胞遗传学研究已经积累了很多，但利用酵母对减数分裂事件进行分子分析的研究才刚刚开始，而在高等生物中的研究还很少。然而，了解和控制减数分裂事件可以为育种，生育和不育问题和人类福利做出不可估量的贡献。本研究利用我们研制的重组抗体，对雄性不育动物牦牛（Bos taurus × Bos grunniens的F_1）、鹿（Cerus elaphus × Cerus nippon F_1）、食蟹猴（M.fascicularis × M.assamesis F_1）的睾丸和卵巢细胞进行了细胞学和分子生物学分析 ...更多信息 A1酶和由减数分裂活性基因制备的各种探针。目前，我们已经完成了对牦牛的研究。昆明研究所的杂交雄性不育鹿还没有拿到手，而杂交雄性不育猴已经死亡，下一步的生产还有待商榷。1.雄性不育主要是由减数分裂降解引起的，在减数分裂终变期和中期Ⅰ也有部分减数分裂降解，但在卵母细胞中也观察到类似的减数分裂降解，表明卵母细胞数量减少。针对RecA蛋白和Lim15制备的抗体（百合减数分裂前期Ⅰ特异基因）蛋白与大多数减数分裂细胞（包括不育睾丸细胞）呈阳性结合，而在体细胞（包括睾丸细胞和卵巢细胞）中几乎没有结合（正常牛）动物基本上是相同的，基于细胞蛋白。3.从不育和可育睾丸减数分裂前期细胞分离的cDNA克隆在限制性图谱和获得的数量上是相似的，表明不育雄性的减数分裂前期细胞中存在正常的转录过程，这为我们推断不育雄性的减数分裂前期细胞中存在正常的转录过程提供了线索。1.所谓雄性不育是由于减数分裂的破坏导致精子减少而引起的。这种情况也发生在女性身上。由于雌性可育性是由一个卵细胞获得的，所以尽管卵母细胞数量减少，雌性仍是可育的。2.迄今为止所研究的分子事件表明，可育性和不育性之间直到前期或中期都没有可检测到的差异。这些杂种小鼠的减数分裂退化可能是由于细胞凋亡引起的，但这需要更多的分析。少

项目成果

Furukawa, K., Inagaki, H., Naruge, T., Tabata, S.A,Hotta, Y., Tomita, T., Yamaguchi, A., Yoshikuni, M.and Nagahama, Y.: "cDNA cloning and functional characterization of meiosis specific protein (MNSI) with apparent nuclear association." Chromosome Researc

Furukawa, K.、Inagaki, H.、Naruge, T.、Tabata, S.A、Hotta, Y.、Tomita, T.、Yamaguchi, A.、Yoshikuni, M. 和 Nagahama, Y.：“cDNA 克隆和功能表征

Sato,S.,Suzuki,H.,Widiastati,U.,Hotta,Y.and Tabata,S.: "Identification and characterization of genes induced during sexual differentiation in Schyzogaccharomyces pombe." Curr.Genetics. 26. 31-37 (1994)

Sato,S.、Suzuki,H.、Widiastati,U.、Hotta,Y.和 Tabata,S.：“粟酒裂殖酵母性分化过程中诱导基因的鉴定和表征。”

Kofman-Alfaro, S., Speed, R.M., Boyle, S.and Chandley, A.C.: "Condensation behavior of the human X chromosome in male germ cells and Sertoli cells examined by fluorescence in situ hybridization." Chromosome Research. 2. 439-444 (1994)

Kofman-Alfaro, S.、Speed, R.M.、Boyle, S. 和 Chandley, A.C.：“通过荧光原位杂交检查男性生殖细胞和支持细胞中人类 X 染色体的凝聚行为。”

Furukawa,K.,Inagaki,H.,Naruge,T.,Tabata,S.,Hotta,Y.,Tomita,T.,Yamaguchi,A.,Yoshikuni,M.and Nagahama,Y.: "cDNA cloning and fanctional characterization of meiosis specific protein(MNSI)with apparentnuclear association" Chromosome Research. 2. 99-113 (1994)

Furukawa,K.、Inagaki,H.、Naruge,T.、Tabata,S.、Hotta,Y.、Tomita,T.、Yamaguchi,A.、Yoshikuni,M. 和 Nagahama,Y.：“cDNA 克隆和功能性

Hotta, Y., Furukawa, K.and Tabata, S.: "Meiosis specific transcription and functional protein." Adv.Biophys.31. 101-115 (1995)

Hotta, Y.、Furukawa, K. 和 Tabata, S.：“减数分裂特异性转录和功能蛋白。”