Expression and immunohistochemical localization of TMEM16A/anoctamin 1, a calcium-activated chloride channel in the mouse cochlea
Expression and immunohistochemical localization of TMEM16A/anoctamin 1, a calcium-activated chloride channel in the mouse cochlea
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DOI：
10.1007/s00441-011-1206-6
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发表时间：
2011-08-01
影响因子：
3.6
通讯作者：
Kim, In-Beom
中科院分区：
文献类型：
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作者：
Jeon, Ji Hyun;Park, Jae Woo;Kim, In-Beom
耳蜗中的声音传导取决于内淋巴中独特的高浓度 K+。高 K+ 浓度的产生和维持伴随着 Cl- 循环。在这项研究中，我们通过蛋白质印迹和免疫组织化学研究了小鼠耳蜗中 TMEM16A/anoctamin 1 (ANO1)（一种最近克隆的 Ca2+ 激活的 Cl-通道）的表达和定位。通过蛋白质印迹法鉴定了耳蜗中的 ANO1 蛋白。在血管纹中发现了一条线，在柯蒂氏器中发现了三个斑块。通过抗密蛋白 11（血管纹基底细胞的标记物）的双标记实验检查了 ANO1 的细胞定位。结果表明，ANO1 与 Claudin 11 共定位，表明其在基底细胞中表达。我们还通过双标记和三标记技术检查了 ANO1 在 Corti 器官中的定位，其中使用抗肌球蛋白 VI（毛细胞标记物）和抗突触素（毛细胞下橄榄耳蜗传出神经末梢标记物）。结果清楚地表明，ANO1 与突触素共定位，但不与肌球蛋白 VI 共定位，表明 ANO1 定位于外毛细胞下方的内侧橄榄耳蜗传出神经末梢。这些结果表明，ANO1 可能专门参与从内侧橄榄耳蜗传出神经末梢到柯蒂氏器外毛细胞的突触传递，以及血管纹基底细胞中的 Cl-循环。
Sound transduction in the cochlea depends on the unique high concentrations of K+ in the endolymph. The production and maintenance of high K+ concentrations are accompanied by Cl- cycling. In this study, we report on an investigation of the expression and localization of TMEM16A/anoctamin 1 (ANO1), a recently cloned Ca2+-activated Cl- channel, in the mouse cochlea by Western blot and immunhistochemistry. The ANO1 protein was identified in the cochlea by Western blotting. The immunoreactivity was found in stria vascularis as a line and in the organ of Corti as three plaques. The cellular localization of ANO1 was examined by means of double-labeling experiments with anti-claudin 11, a marker for basal cells of the stria vascularis. The results demonstrated that ANO1 colocalized with claudin 11, indicating its expression in basal cells. We also examined ANO1 localization in the organ of Corti by double- and triple-labeling techniques with anti-myosin VI, a marker for hair cells, and anti-synaptophysin, a marker for olivocochlear efferent nerve endings under hair cells. The results clearly showed that ANO1 is colocalized with synaptophysin, but not with myosin VI, indicating that ANO1 is localized at medial olivocochlear efferent nerve endings under outer hair cells. These results suggest that ANO1 may be specifically involved in synaptic transmission from medial olivocochlear efferent nerve endings to outer hair cells in the organ of Corti, as well as Cl- cycling in basal cells of the stria vascularis.