Expression of prestin in OHCs is reduced in Spag6 gene knockout mice.
Expression of prestin in OHCs is reduced in Spag6 gene knockout mice.
复制标题
Spag6 基因敲除小鼠中 OHC 中 prestin 的表达降低。
DOI：
10.1016/j.neulet.2015.03.007
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发表时间：
2015-04-10
影响因子：
2.5
通讯作者：
Li J
中科院分区：
文献类型：
--
作者：
Wang J;Li X;Zhang Z;Wang H;Li J
精子相关抗原6（Spag 6）基因编码一种轴丝蛋白（SPAG 6），广泛表达于含有纤毛细胞的组织和器官中。本研究旨在探讨SPAG 6是否在耳蜗毛细胞中表达，以及普雷斯廷与SPAG 6之间可能存在的相关性。免疫荧光染色观察SPAG 6在Corti器中的分布及基底膜毛细胞的形态学特征。分别用Western blotting和real-time PCR检测Spag 6突变小鼠中普雷斯廷的含量。此外，进行免疫共沉淀试验以证实推测的普雷斯廷和SPAG 6之间的相互作用。我们观察到SPAG 6在外毛细胞（OHC）的表皮板中表达，普雷斯廷在OHC的侧壁中与SPAG 6沿着定位在该位点。与Spag 6 +/+小鼠相比，Spag 6 −/−小鼠表现出明显的OHC形态学异常和较低的普雷斯廷荧光强度。Spag 6 −/−小鼠中普雷斯廷的表达在蛋白和mRNA水平上均显著降低。此外，免疫共沉淀试验证明了普雷斯廷和SPAG 6之间的相互作用。综上所述，这些数据表明，SPAG 6通过维持普雷斯廷的正常表达而对OHC的稳定性是不可或缺的，这意味着Spag 6基因对于OHC的机械感觉功能是必需的。
Sperm-associated antigen 6 (Spag6) gene, which encodes an axonemal protein (SPAG6), ubiquitously expresses in tissue and organs containing ciliated cells. The present work was to investigate whether SPAG6 expressed in cochlear hair cells and, if so, to explore the presumable correlations between prestin and SPAG6. The distribution of SPAG6 in organ of Corti and the morphological features of hair cells in basilar membrane were investigated by immunofluorescent staining. The amount of prestin in Spag6 mutant mice was measured by Western blotting and real-time PCR, respectively. Additionally, co-immunoprecipitation tests were performed to confirm the presumed interaction between prestin and SPAG6. We observed that SPAG6 expressed in the cuticular plate in outer hair cells (OHCs) and prestin in the lateral wall of OHCs that located along with SPAG6 at this site. In comparison to Spag6 +/+ mice, Spag6 −/− mice showed apparent morphological abnormity of OHCs and lower intensity of prestin fluorescence. The expression of prestin in Spag6 −/− mice reduced significantly at both protein and mRNA levels. Moreover, co-immunoprecipitation tests demonstrated the interaction between prestin and SPAG6. Taken together, these data indicate that SPAG6 is indispensible for the stability of OHCs by maintaining the normal expression of prestin, which implies that Spag6 gene is essential for mechanosensory function of OHCs.