BMP4和SMAD4基因在公绵羊繁殖相关组织的表达
|
摘要
试验旨在探索骨形态发生蛋白4(Bone morphogenetic protein 4,BMP4)和SMAD家族蛋白4(SMAD family member 4,SMAD4)基因在公绵羊繁殖中的作用,以便深入研究公绵羊繁殖机制。采用荧光定量PCR技术检测BMP4和SMAD4基因在高繁殖力的小尾寒羊公羊(n=3)和低繁殖力苏尼特公羊(n=3)大脑、小脑、下丘脑、垂体、输精管、肾上腺、睾丸和附睾8种组织中的表达。结果表明:BMP4和SMAD4基因在小尾寒羊公羊和苏尼特羊公羊各种组织中均有表达。其中BMP4基因在睾丸中高表达,在输精管、肾上腺中中等表达,在其他组织中均呈痕量表达;SMAD4基因在睾丸、下丘脑、小脑、大脑中高表达,在其他组织中中等表达。BMP4基因在低繁殖力苏尼特羊公羊睾丸中表达量显著高于高繁殖力小尾寒羊公羊(P<0.05);SMAD4基因在低繁殖力苏尼特公羊8种组织中表达量均显著高于高繁殖力小尾寒羊公羊(P<0.05)。说明BMP4和SMAD4基因在公绵羊繁殖中起到一定程度的负调控作用,这为公羊高繁殖性状选育提供了参考依据。
To reveal the roles of bone morphogenetic protein 4(BMP4)and SMAD family member 4(SMAD4) genes in ram reproduction and research the mechanism of prolificacy in rams. Real-time fluorescence quantitative PCR(RT-q PCR) was performed to detect BMP4 and SMAD4 m RNA expression in brain, cerebellum, hypothalamus, pituitary, vas deferens, adrenal gland, testis, epididymis of multiparous Small-tailed han sheep(n=3)and uniparous Sunite sheep(n=3).The results showed that both BMP4 and SMAD4 genes were expressed in eight tissues of rams. The expression of BMP4 was high in testis and moderate in the vas deferens and adrenal gland, meanwhile expression in other tissues was low. The expression of SMAD4 was high in testis,hypothalamus, cerebellum, brain and moderate in other tissues. The expression of BMP4 in testis in the uniparous Sunite was significantly higher than that in multiparous Small-tailed han sheep(P<0.05). The expression of SMAD4 in eight tissues in uniparous Sunite was significantly higher than that in multiparous Small-tailed han sheep(P<0.05). The results suggest that BMP4 and SMAD4 genes may play a negative role in the regulation of ram reproduction, which provides a reference for the breeding of high reproductive traits in male sheep.
To reveal the roles of bone morphogenetic protein 4(BMP4)and SMAD family member 4(SMAD4) genes in ram reproduction and research the mechanism of prolificacy in rams. Real-time fluorescence quantitative PCR(RT-q PCR) was performed to detect BMP4 and SMAD4 m RNA expression in brain, cerebellum, hypothalamus, pituitary, vas deferens, adrenal gland, testis, epididymis of multiparous Small-tailed han sheep(n=3)and uniparous Sunite sheep(n=3).The results showed that both BMP4 and SMAD4 genes were expressed in eight tissues of rams. The expression of BMP4 was high in testis and moderate in the vas deferens and adrenal gland, meanwhile expression in other tissues was low. The expression of SMAD4 was high in testis,hypothalamus, cerebellum, brain and moderate in other tissues. The expression of BMP4 in testis in the uniparous Sunite was significantly higher than that in multiparous Small-tailed han sheep(P<0.05). The expression of SMAD4 in eight tissues in uniparous Sunite was significantly higher than that in multiparous Small-tailed han sheep(P<0.05). The results suggest that BMP4 and SMAD4 genes may play a negative role in the regulation of ram reproduction, which provides a reference for the breeding of high reproductive traits in male sheep.
引文
[1]董莲花,冉茂良,李智,等.TGF-β超家族信号通路及其调控睾丸发育和精子生成的研究进展[J].中国畜牧杂志,2016,52(3):86-92.
[2]李毅.Bmp4/Smad信号通路在小鼠精原干细胞分化过程中的作用及其机制研究[D].济南:山东大学,2012.
[3]Chen S R,Liu Y X.Regulation of spermatogonial stem cell selfrenewal and spermatocyte meiosis by sertoli[J].Reproduction,2015,36(6):489-495.
[4]Chen S R,Tang J X,Cheng J M,et al.Loss of Gata4 in Sertoli cells impairs the spermatogonial stem cell niche and causes germ cell exhaustion by attenuating chemokine signaling[J].Oncotarget,2015,6(35):37012-37027.
[5]Archambeault D R,Yao H C.Loss of smad4 in Sertoli and Leydig cells leads to testicular dysgenesis and hemorrhagic tumor formation in mice[J].Biology of Reproduction,2014,90(3):62.
[6]周梅,曹晓涵,贺小云,等.绵羊FGF 7基因组织表达及其多态性与产羔数之间的关系[J].畜牧兽医学报,2018,49(3):525-533.
[7]Schmittgen T D,Livak K J.Analyzing real-time PCR data by the comparative CT method[J].Nature Protocols,2008,3(6):1101.
[8]Bier E,De Robertis E M.BMP gradients:A paradigm for morphogen-mediated developmental patterning[J].Science,2015,348(6242):5838.
[9]Hai Y,Sun M,Niu M,et al.BMP4 promotes human sertoli cell proliferation via Smad1/5 and ID2/3 pathway and its abnormality is associated with azoospermia[J].Discovery Medicine,2015,19(105):311-325.
[10]Tan S,Feng B,Yin M,et al.Stromal Senp1 promotes mouse early folliculogenesis by regulating BMP4 expression[J].Cell&Bioscience,2017,7(1):36-36.
[11]Da C E,De Souza G B,Passos J R,et al.Effects of bone morphogenetic protein 4(BMP4)on in vitro development and survival of bovine preantral follicles enclosed in fragments ovarian tissue[J].Zygote,2017,25(3):256-264.
[12]徐业芬.Smad信号转导通路相关基因表达与湖羊高繁殖力关系的研究[D].南京:南京农业大学,2010.
[13]孙振梅.黔北麻羊BMP4和BMPR-IB基因克隆表达及其多态性与繁殖性状的关联分析[D].贵阳:贵州大学,2017.
[14]秦立红,张国梁,吴健,等.BMP2和BMP4基因在草原红牛睾丸组织中的表达量分析[J].中国兽医学报,2016,36(4):661-664.
[15]Macias M J,Martin-Malpartida P,MassaguéJ.Structural determinants of Smad function in TGF-βsignaling[J].Trends in Biochemical Sciences,2015,40(6):296-308.
[16]王玉恒,徐业芬,牛家强,等.牦牛Smad 4基因m RNA组织表达谱及其mi RNAs初步研究[J].畜牧兽医学报,2017,48(10):1882-1891.
[17]郝晓霞.雄性生殖细胞SMAD4在小鼠精子发生及AR在人睾丸生殖细胞瘤中的功能研究[D].北京:中国科学院大学,2017.
[18]高真真.生长分化因子9及其信号通路蛋白(ALK5、Smad2和Smad4)在成年鹌鹑和鸡睾丸中的表达与定位[D].太谷:山西农业大学,2014.
[2]李毅.Bmp4/Smad信号通路在小鼠精原干细胞分化过程中的作用及其机制研究[D].济南:山东大学,2012.
[3]Chen S R,Liu Y X.Regulation of spermatogonial stem cell selfrenewal and spermatocyte meiosis by sertoli[J].Reproduction,2015,36(6):489-495.
[4]Chen S R,Tang J X,Cheng J M,et al.Loss of Gata4 in Sertoli cells impairs the spermatogonial stem cell niche and causes germ cell exhaustion by attenuating chemokine signaling[J].Oncotarget,2015,6(35):37012-37027.
[5]Archambeault D R,Yao H C.Loss of smad4 in Sertoli and Leydig cells leads to testicular dysgenesis and hemorrhagic tumor formation in mice[J].Biology of Reproduction,2014,90(3):62.
[6]周梅,曹晓涵,贺小云,等.绵羊FGF 7基因组织表达及其多态性与产羔数之间的关系[J].畜牧兽医学报,2018,49(3):525-533.
[7]Schmittgen T D,Livak K J.Analyzing real-time PCR data by the comparative CT method[J].Nature Protocols,2008,3(6):1101.
[8]Bier E,De Robertis E M.BMP gradients:A paradigm for morphogen-mediated developmental patterning[J].Science,2015,348(6242):5838.
[9]Hai Y,Sun M,Niu M,et al.BMP4 promotes human sertoli cell proliferation via Smad1/5 and ID2/3 pathway and its abnormality is associated with azoospermia[J].Discovery Medicine,2015,19(105):311-325.
[10]Tan S,Feng B,Yin M,et al.Stromal Senp1 promotes mouse early folliculogenesis by regulating BMP4 expression[J].Cell&Bioscience,2017,7(1):36-36.
[11]Da C E,De Souza G B,Passos J R,et al.Effects of bone morphogenetic protein 4(BMP4)on in vitro development and survival of bovine preantral follicles enclosed in fragments ovarian tissue[J].Zygote,2017,25(3):256-264.
[12]徐业芬.Smad信号转导通路相关基因表达与湖羊高繁殖力关系的研究[D].南京:南京农业大学,2010.
[13]孙振梅.黔北麻羊BMP4和BMPR-IB基因克隆表达及其多态性与繁殖性状的关联分析[D].贵阳:贵州大学,2017.
[14]秦立红,张国梁,吴健,等.BMP2和BMP4基因在草原红牛睾丸组织中的表达量分析[J].中国兽医学报,2016,36(4):661-664.
[15]Macias M J,Martin-Malpartida P,MassaguéJ.Structural determinants of Smad function in TGF-βsignaling[J].Trends in Biochemical Sciences,2015,40(6):296-308.
[16]王玉恒,徐业芬,牛家强,等.牦牛Smad 4基因m RNA组织表达谱及其mi RNAs初步研究[J].畜牧兽医学报,2017,48(10):1882-1891.
[17]郝晓霞.雄性生殖细胞SMAD4在小鼠精子发生及AR在人睾丸生殖细胞瘤中的功能研究[D].北京:中国科学院大学,2017.
[18]高真真.生长分化因子9及其信号通路蛋白(ALK5、Smad2和Smad4)在成年鹌鹑和鸡睾丸中的表达与定位[D].太谷:山西农业大学,2014.