PRDM6属于PRDM蛋白家族的一员,PRDM6基因编码1个转录抑制因子,具有与DNA、RNA、蛋白质结合的能力,同时还具有组蛋白赖氨酸N-甲基转移酶活性。全基因组关联分析结果显示,PRDM6基因遗传变异与骨密度和体质量等性状显著相关,且该基因插入/...PRDM6属于PRDM蛋白家族的一员,PRDM6基因编码1个转录抑制因子,具有与DNA、RNA、蛋白质结合的能力,同时还具有组蛋白赖氨酸N-甲基转移酶活性。全基因组关联分析结果显示,PRDM6基因遗传变异与骨密度和体质量等性状显著相关,且该基因插入/缺失(Indel)遗传变异还被发现与陕北白绒山羊的多个生长性状显著相关。因此,本研究将PRDM6基因作为研究对象,以期挖掘到与内蒙古绒山羊生长性状相关的关键遗传变异位点。结合Ensembl数据库提供的山羊PRDM6遗传变异信息和已发表文章中的遗传变异位点,本研究探究了2个InDel位点(rs656578433、rs651603667)多态性及其在内蒙古绒山羊群体中的分布规律。发现在本研究检测的群体中,只有第一内含子区的12 bp InDel(rs651603667)存在多态性。不同基因型与生长性状的关联分析结果显示,rs651603667存在3种基因型,即纯合插入型(II)、杂合型(ID)和纯合缺失型(DD),频率分别为0.399、0.444、0.159。在育成羊群体中,此rs651603667突变与体质量、胸深显著相关(P<0.05);在成年羊群体中,rs651603667突变与体长、髋宽显著相关(P<0.05),和体高、胸深极极显著相关(P<0.01);此外,皮尔逊(Pearson)相关分析结果显示,该突变位点显著影响的生长性状均与体质量存在极显著相关(P<0.01)。综上,PRDM6基因12 bp InDel突变位点与内蒙古绒山羊体长、髋宽和体高、胸深具有显著或极显著相关性,可作为内蒙古绒山羊生长性状选育的有效分子标记。展开更多
Studies have demonstrated that regulation of GDNF on male germline stem cells （mGSCs） mainly through Ras/Erk1/2, Src family kinase and PI3K/Akt signaling pathways, but the signaling pathways GDNF-mediated are differ...Studies have demonstrated that regulation of GDNF on male germline stem cells （mGSCs） mainly through Ras/Erk1/2, Src family kinase and PI3K/Akt signaling pathways, but the signaling pathways GDNF-mediated are different when the species and cell lines varied. Whether GDNF regulates self-renewal of mGSCs isolated from livestock has not been reported. Here, we purified mGSCs from dairy goat testis using mixed enzymes and fibronectin. Immunofluoresce staining revealed the cultured dairy mGSCs expressed Vasa, Nanos2, Ngn3, Tert, Dazl, Lin28, Oct4, CD49f, Stra8 and GFRa1, reflecting that these cells were mGSCs phenotype. Then we cultured these dairy goat mGSCs in different concentrations of GDNF （0, 5, 10, or 20 ng mL-1） to optimize the best concentration of GDNF to sustain the dairy goat mGSCs self-renewal, after that the inhibitor of PI3K （LY294002, 10 μmol L-1） was added to the medium which contains the optimal concentration of GDNF we obtained by experiments. The mGSCs cultured in different media were compared through the population doubling time （PDT）, capacity of cell proliferation evaluated by PCNA and BrdU immunofluorescence staining, RT-PCR, QRT-PCR, Western blotting and flow cytometry. Results showed that 10 ng mL-1 was the optimal concentration of GDNF to maintain goat mGSCs self-renewal and GDNF up-regulates c-Myc transcription via the PI3K/Akt pathway to promote goat mGSCs proliferation. This study provides us an efficient model to study the mechanism in mGSCs proliferation and differentiation in goat, and has important implications in unveiling signaling pathways in livestock GSCs.展开更多
Adhesions between different cells and extracellular matrix have been studied extensively in vitro, but little is known about their functions in testicular tissue counterparts. Spermatogonia and their companion somatic...Adhesions between different cells and extracellular matrix have been studied extensively in vitro, but little is known about their functions in testicular tissue counterparts. Spermatogonia and their companion somatic cells maintain a close association throughout spermatogenesis and this association is necessary for normal spermatogenesis. In order to keep the relative integrity of the testicular tissues, and to detect the development in vitro, culture testicular tissues in a three- dimensional （3D） agarose matrix was examined. Testicular tissues isolated from 6.5 d postpartum （dpp） mouse were cultured on the top of the matrix for 26 d with a medium height up to 4/5 of the 3D agarose matrix. The results showed that in this 3D culture environment, each type of testicular cells kept the same structure, localization and function as in vivo and might be more biologically relevant to living organisms. After culture, germ cell marker VASA and meiosis markers DAZL and SCP3 showed typical positive analysed by immunofluorescence staining and RT-PCR. It demonstrated that this 3D culture system was able to maintain the number of germ cells and promote the meiosis initiation of male germ cells.展开更多
文摘PRDM6属于PRDM蛋白家族的一员,PRDM6基因编码1个转录抑制因子,具有与DNA、RNA、蛋白质结合的能力,同时还具有组蛋白赖氨酸N-甲基转移酶活性。全基因组关联分析结果显示,PRDM6基因遗传变异与骨密度和体质量等性状显著相关,且该基因插入/缺失(Indel)遗传变异还被发现与陕北白绒山羊的多个生长性状显著相关。因此,本研究将PRDM6基因作为研究对象,以期挖掘到与内蒙古绒山羊生长性状相关的关键遗传变异位点。结合Ensembl数据库提供的山羊PRDM6遗传变异信息和已发表文章中的遗传变异位点,本研究探究了2个InDel位点(rs656578433、rs651603667)多态性及其在内蒙古绒山羊群体中的分布规律。发现在本研究检测的群体中,只有第一内含子区的12 bp InDel(rs651603667)存在多态性。不同基因型与生长性状的关联分析结果显示,rs651603667存在3种基因型,即纯合插入型(II)、杂合型(ID)和纯合缺失型(DD),频率分别为0.399、0.444、0.159。在育成羊群体中,此rs651603667突变与体质量、胸深显著相关(P<0.05);在成年羊群体中,rs651603667突变与体长、髋宽显著相关(P<0.05),和体高、胸深极极显著相关(P<0.01);此外,皮尔逊(Pearson)相关分析结果显示,该突变位点显著影响的生长性状均与体质量存在极显著相关(P<0.01)。综上,PRDM6基因12 bp InDel突变位点与内蒙古绒山羊体长、髋宽和体高、胸深具有显著或极显著相关性,可作为内蒙古绒山羊生长性状选育的有效分子标记。
基金supported by the National Natural Science Foundation of China(30972097,31272518)the Program for the New Century Excellent Talents of State Ministry of Education of P.R.China(NCET-09-0654)+2 种基金the Doctoral Fund of Ministry of Education of P.R.China(RFDP,20120204110030)the Scientific Research Program of Shaanxi Province,China(2011K02-06)the Fundamental Research Funds for the Central Universities,China(QN2011012)
文摘Studies have demonstrated that regulation of GDNF on male germline stem cells （mGSCs） mainly through Ras/Erk1/2, Src family kinase and PI3K/Akt signaling pathways, but the signaling pathways GDNF-mediated are different when the species and cell lines varied. Whether GDNF regulates self-renewal of mGSCs isolated from livestock has not been reported. Here, we purified mGSCs from dairy goat testis using mixed enzymes and fibronectin. Immunofluoresce staining revealed the cultured dairy mGSCs expressed Vasa, Nanos2, Ngn3, Tert, Dazl, Lin28, Oct4, CD49f, Stra8 and GFRa1, reflecting that these cells were mGSCs phenotype. Then we cultured these dairy goat mGSCs in different concentrations of GDNF （0, 5, 10, or 20 ng mL-1） to optimize the best concentration of GDNF to sustain the dairy goat mGSCs self-renewal, after that the inhibitor of PI3K （LY294002, 10 μmol L-1） was added to the medium which contains the optimal concentration of GDNF we obtained by experiments. The mGSCs cultured in different media were compared through the population doubling time （PDT）, capacity of cell proliferation evaluated by PCNA and BrdU immunofluorescence staining, RT-PCR, QRT-PCR, Western blotting and flow cytometry. Results showed that 10 ng mL-1 was the optimal concentration of GDNF to maintain goat mGSCs self-renewal and GDNF up-regulates c-Myc transcription via the PI3K/Akt pathway to promote goat mGSCs proliferation. This study provides us an efficient model to study the mechanism in mGSCs proliferation and differentiation in goat, and has important implications in unveiling signaling pathways in livestock GSCs.
基金supported by the National Natural Science Foundation of China(31272518)the program for the New Century Excellent Talents of Ministry of Education of China(NCET-09-0654)+1 种基金the Doctoral Fund of Ministry of Education of P.R.China(RFDP,20120204110030)the Fundamental Research Funds for the Central Universities,China(QN2011012)
文摘Adhesions between different cells and extracellular matrix have been studied extensively in vitro, but little is known about their functions in testicular tissue counterparts. Spermatogonia and their companion somatic cells maintain a close association throughout spermatogenesis and this association is necessary for normal spermatogenesis. In order to keep the relative integrity of the testicular tissues, and to detect the development in vitro, culture testicular tissues in a three- dimensional （3D） agarose matrix was examined. Testicular tissues isolated from 6.5 d postpartum （dpp） mouse were cultured on the top of the matrix for 26 d with a medium height up to 4/5 of the 3D agarose matrix. The results showed that in this 3D culture environment, each type of testicular cells kept the same structure, localization and function as in vivo and might be more biologically relevant to living organisms. After culture, germ cell marker VASA and meiosis markers DAZL and SCP3 showed typical positive analysed by immunofluorescence staining and RT-PCR. It demonstrated that this 3D culture system was able to maintain the number of germ cells and promote the meiosis initiation of male germ cells.
