Department of Anatomy & Cell Biology, Wayne State University School of Medicine, Detroit, MI 48201, United States.
Neuroscience. 2012 Nov 8;224:63-9. doi: 10.1016/j.neuroscience.2012.08.017. Epub 2012 Aug 19.
Retinal bipolar cells and ganglion cells are known to possess voltage-gated T-type Ca(2+) channels. Previous electrophysiological recording studies suggested that there is differential expression of different T-type Ca(2+) channel α1 subunits among bipolar cells. The detailed expression patterns of the individual T-type Ca(2+) channel subunits in the retina, however, remain unknown. In this study, we examined the expression of the Ca(V)3.2 Ca(2+) channel α1 subunit in the mouse retina using immunohistochemical analysis and patch-clamp recordings together with a Ca(V)3.2 knock out (KO) mouse line. The specificity of a Ca(V)3.2 Ca(2+) channel antibody was first confirmed in recombinant T-type Ca(2+) channels expressed in human embryonic kidney (HEK) cells and in Ca(V)3.2 KO mice. Our immunohistochemical analysis indicates that the Ca(V)3.2 antibody labels a subgroup of type-3 cone bipolar cells (CBCs), the PKAβII-immunopositive type-3 CBCs. The labeling was observed throughout the cell including dendrites and axon terminals. Our patch-clamp recording results further demonstrate that Ca(V)3.2 Ca(2+) channels contribute to the T-type Ca(2+) current in a subpopulation of type-3 CBCs. The findings of this study provide new insights into understanding the functional roles of T-type Ca(2+) channels in retinal processing.
已知视网膜双极细胞和节细胞具有电压门控 T 型 Ca(2+)通道。先前的电生理记录研究表明,双极细胞中存在不同 T 型 Ca(2+)通道 α1 亚基的差异表达。然而,视网膜中单个 T 型 Ca(2+)通道亚基的详细表达模式仍不清楚。在这项研究中,我们使用免疫组织化学分析和膜片钳记录以及 Ca(V)3.2 敲除 (KO) 小鼠系来检查 Ca(V)3.2 Ca(2+)通道 α1 亚基在小鼠视网膜中的表达。首先在重组 T 型 Ca(2+)通道中证实了 Ca(V)3.2 Ca(2+)通道抗体的特异性,该通道在人胚肾 (HEK) 细胞和 Ca(V)3.2 KO 小鼠中表达。我们的免疫组织化学分析表明,Ca(V)3.2 抗体标记了一小群 3 型锥形双极细胞 (CBCs),即 PKAβII 免疫阳性的 3 型 CBCs。该标记在整个细胞中都有观察到,包括树突和轴突末梢。我们的膜片钳记录结果进一步表明,Ca(V)3.2 Ca(2+)通道有助于 3 型 CBC 亚群中的 T 型 Ca(2+)电流。这项研究的发现为理解 T 型 Ca(2+)通道在视网膜处理中的功能作用提供了新的见解。