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隐睾小鼠与正常成年小鼠睾丸蛋白表达谱的差异分析
  • ISSN号:0371-0874
  • 期刊名称:《生理学报》
  • 时间:0
  • 分类:Q51[生物学—生物化学]
  • 作者机构:[1]军事医学科学院基础医学研究所,北京100850, [2]黄淮学院农林科学系,驻马店463000, [3]吉林大学畜牧兽医学院基础兽医系,长春130002
  • 相关基金:This work was supported by the National Natural Science Foundation of China (No. 30471246).
中文摘要:

为使精原干细胞(spermatogonial stem cells,SSCs)在体外大量扩增,需要阐明SSCs自增殖机制。为筛选SSCs自增殖相关因子,探索SSCs自增殖机制,本研究选取10日龄昆明乳鼠行隐睾手术,术后35d分别取小鼠两侧睾丸。组织学分析结果显示,实验性隐睾中生殖细胞的分化停滞在精母细胞阶段,且只有少量的精母细胞出现,精原细胞的比例高于正常成年雄性小鼠(45日龄)。应用双向凝胶电泳分析隐睾小鼠与正常成年小鼠睾丸差异表达蛋白。结果显示,与正常成年小鼠相比,隐睾小鼠睾丸中有9种蛋白表达发生了显著变化,其中6种蛋白表达下调,3种上调。对9种差异表达蛋白点胶内酶切后进行质谱分析,其中4种蛋白分别鉴定为磷脂酰乙醇胺结合蛋白1(phosphatidylethanolamine-binding protein1,PEBP1),HES—related basic helix-100p-helix protein(HERP),Stathmin蛋白和一种未命名蛋白。本研究通过制作有效的隐睾动物模型,运用蛋白组学的技术方法,成功筛选并鉴定了4种隐睾相关蛋白,有助于探讨SSCs自增殖及隐睾引起雄性不育的机制。

英文摘要:

To screen factors related to spermatogonial stem cell (SSC) proliferation, and to investigate the mechanism of infertility caused by cryptorchidism, ten-day-old Kunming (KM) mice were used and experimental cryptorchidism was conducted. On the 35th day after cryptorchid operation, the left testes were fixed in Bouin's fluid and used for histological analysis. The testes of 45-day-old mice were subjected to the same histological analysis, and it was found that they contained germ cells at every stage of development, from SSCs to sperm, indicating that the animals were fully sexually mature at this age. While in experimental cryptorchid mice, the spermatogenesis was arrested at the stage of spermatocytes, and only spermatogonia and primary spermatocytes were present in cryptorchid testes. The proportion of spermatogonia to other types of germ cells was much higher than that in sexually mature mice. On the other hand, the fight testes were used for proteomic analysis. The total protein in testes was extracted on the 35th day after cryptorchid operation. The differentially expressed proteins in cryptorchid mice and sexually mature mice were screened and compared by the proteomic techniques. Through the separation of two-dimensional gel electrophoresis (2-DE), 20 differential protein spots were found, and 9 of them were digested and identified by the matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectrum. In cryptorchid mice, 6 out of 9 proteins were down-regulated, and 3 were up-regulated. Among these proteins, 4 proteins were identified, and they were Stathmin, phosphatidylethanolamine-binding protein1 (PEBP1), HES-related basic helix-loop-helixprotein (HERP), and one unnamed protein (we temporarily named it Px). More Stathmin, PEBP 1 and Px were expressed in sexually mature mice than in experimental cryptorchid mice. But HERP1 was the other way round. In the present study, we have screened 4 proteins related to cryptorchidism. It is helpful to study the mec

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期刊信息
  • 《生理学报》
  • 中国科技核心期刊
  • 主管单位:中国科学院
  • 主办单位:中国生理学会 中国科学院上海生理研究所
  • 主编:王建军
  • 地址:上海岳阳路319号31B楼
  • 邮编:200031
  • 邮箱:actaps@sibs.ac.cn
  • 电话:021-54922832
  • 国际标准刊号:ISSN:0371-0874
  • 国内统一刊号:ISSN:31-1352/Q
  • 邮发代号:4-157
  • 获奖情况:
  • 中国自然科学核心期刊,中国科学院优秀期刊特别奖,荣获首届国家期刊奖
  • 国内外数据库收录:
  • 美国化学文摘(网络版),波兰哥白尼索引,荷兰文摘与引文数据库,美国生物医学检索系统,美国生物科学数据库,日本日本科学技术振兴机构数据库,中国中国科技核心期刊,中国北大核心期刊(2004版),中国北大核心期刊(2008版),中国北大核心期刊(2011版),中国北大核心期刊(2014版),中国北大核心期刊(2000版)
  • 被引量:8098