Prestin在Spag6基因敲除小鼠外毛细胞中的表达

发布时间：2018-03-14 10:37

本文选题：精子相关抗原　切入点：Prestin　出处：《山东大学学报(医学版)》2015年08期 　论文类型：期刊论文

【摘要】：目的探究prestin在精子相关抗原6(Spag6)基因敲除小鼠中的表达及其与SPAG6之间的关系。方法获取小鼠耳蜗基底膜组织,进行免疫染色,采用共聚焦显微镜观察prestin与SPAG6在毛细胞中的共同分布。比较野生小鼠(Spag6+/+)、杂合型小鼠(Spag6+/-)和同窝基因敲除小鼠(Spag6-/-)外毛细胞prestin的荧光强度差异和毛细胞的形态改变,并以蛋白印记实验和实时定量聚合酶链反应检测3种基因型小鼠的prestin表达水平。结果SPAG6存在于野生型和杂合型成年小鼠的外毛细胞中,在表皮板处呈阳性表达,与prestin共同分布于外毛细胞的外侧壁处。基因敲除小鼠prestin的荧光强度较野生小鼠更弱,并且外毛细胞出现形态异常。蛋白印迹实验和实时定量聚合酶链反应均显示,基因敲除小鼠prestin表达量显著降低。结论SPAG6参与外毛细胞的构成,且影响外毛细胞prestin表达,提示两种蛋白之间存在相互作用。
[Abstract]:Objective to investigate the expression of prestin and its relationship with SPAG6 in spermatozoa associated antigen 6 Spag6 gene knockout mice. The co-distribution of prestin and SPAG6 in hair cells was observed by confocal microscopy. The fluorescence intensity of prestin and the morphological changes of hair cells in wild mice, heterozygous mice, heterozygous mice and in the same nest gene knockout mice were compared. Protein imprinting assay and real-time quantitative polymerase chain reaction were used to detect the expression of prestin in three genotypes of mice. Results SPAG6 was found in the outer hair cells of wild and heterozygous adult mice and expressed positively in the epidermis. The fluorescence intensity of prestin in knockout mice was weaker than that of wild mice, and the morphology of outer hair cells was abnormal. Western blot assay and real-time quantitative polymerase chain reaction showed that the fluorescence intensity of prestin in knockout mice was lower than that in wild mice. Conclusion SPAG6 is involved in the formation of outer hair cells and affects the expression of prestin in outer hair cells, suggesting that there is an interaction between the two proteins.
【作者单位】：山东大学附属省立医院耳鼻咽喉-头颈外科;山东大学医学院病理学与病理生理学教研室;山东省耳科学重点实验室;宾夕法尼亚大学生殖医学及女性健康研究中心;
【基金】：国家自然科学基金(803300121,81072200) 山东省自然科学基金(ZR2006GG2202)
【分类号】：R764

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