摘要
研究TGF-β1在羊驼睾丸的表达与定位,探索TGF-β1在羊驼睾丸发育及精子发生中的作用。以12和24月龄的羊驼睾丸为研究对象,采用Western blotting技术,对TGF-β1在羊驼睾丸蛋白水平的表达进行研究;采用免疫组织化学SABC法对TGF-β1在羊驼睾丸的表达进行组织学定位。Western blotting结果显示羊驼睾丸组织粗蛋白提取物中存在分子量约为25 ku且与多克隆兔抗TGF-β1抗体发生免疫阳性反应的蛋白条带;免疫组化结果显示TGF-β1在12和24月龄羊驼睾丸的间质细胞、支持细胞和各级生精细胞均有不同程度的表达。光密度测定结果显示:TGF-β1在12、24月龄羊驼睾丸间质细胞的光密度分别为0.730 5±0.011 5、0.562 6±0.012 2;在支持细胞分别为0.420 7±0.013 0、0.325 7±0.008 2;在精原细胞分别为0.457 9±0.009 2、0.374 3±0.015 8(P<0.01);在管周肌样细胞分别为0.645 5±0.082 0、0.656 9±0.023 0(P>0.05);TGF-β1在同一年龄阶段不同细胞的表达差异显著(P<0.01,P<0.05)。结果表明羊驼睾丸有TGF-β1的表达;TGF-β1参与羊驼睾丸发育与精子发生的调控。
Transforming growth factor-β1（TGF-β1）plays important roles during testicular development and spermatogenesis,the main aim of the study is to determine the expression and localization of TGF-β1 in alpaca testis.The testes were obtained from alpacas at the age of 12-and 24-months（n=3）at the Scientific Research Base of Shanxi Agricultural University.The protein expression of TGF-β1 in alpaca testes was examined by Western blotting.The cellular localization and developmental changes were examined by immunohistochemistry SABC.The immunostaining of TGF-β1 was present in leydig cells,sertoli cells,myoid cells and spermatogenic cells in the testis of alpacas at the age of 12-and 24-months,the optical density（OD）of TGF-β1 were respectively 0.730 5±0.011 5 and 0.562 6±0.012 2 in leydig cell;0.420 7±0.013 0 and 0.325 7±0.008 2 in sertoli cells;0.645 5±0.082 0 and 0.656 9±0.023 0 in myoid cells;0.457 9±0.009 2 and 0.374 3±0.015 8 in spermatogonia（P〈0.01）.The positive immunoreactivity was located at the cytoplasm,not in the nucleus.No positive immunoreactivity was observed in the negative control.From the changes in pattern of localization of this peptide,we conclude that TGF-β1 is strictly regulated in leydig cells,sertoli cells and germ cells.It suggests that TGF-β1 involves in the regulation of spermatogenesis,which provides direct evidences for TGF-β1 action in the alpaca testis during postnatal development and spermatogenesis.
出处
《畜牧兽医学报》
CAS
CSCD
北大核心
2008年第1期97-102,共6页
ACTA VETERINARIA ET ZOOTECHNICA SINICA
基金
引进国际先进农业科学项目(948项目)(202096)(2003-Z86)资助
