Kawasaki Fumiko, Zou Beiyan, Xu Xia, Ordway Richard W
Department of Biology and Genetics Graduate Program, Pennsylvania State University, University Park, Pennsylvania 16802, USA.
J Neurosci. 2004 Jan 7;24(1):282-5. doi: 10.1523/JNEUROSCI.3553-03.2004.
Presynaptic calcium channels play a central role in chemical synaptic transmission by providing the calcium trigger for evoked neurotransmitter release. These voltage-gated calcium channels are composed of a primary structural subunit, alpha1, as well as auxiliary beta and alpha2delta subunits. Our previous genetic, molecular, and functional analysis has shown that the cacophony (cac) gene encodes a primary presynaptic calcium channel alpha1 subunit in Drosophila. Here we report that transgenic expression of a cac-encoded alpha1 subunit fused with enhanced green fluorescent protein efficiently rescues cac lethal mutations and allows in vivo analysis of calcium channel localization at active zones. The results reported here further characterize the primary role of cac-encoded calcium channels in neurotransmitter release. In addition, these studies provide a unique genetic tool for live imaging of functional presynaptic calcium channels in vivo and define a molecular marker for immunolocalization of other presynaptic proteins relative to active zones. These findings are expected to facilitate additional analysis of synaptic development and function in this important model system.
突触前钙通道通过为诱发的神经递质释放提供钙触发信号,在化学突触传递中发挥核心作用。这些电压门控钙通道由一个主要结构亚基α1以及辅助β和α2δ亚基组成。我们之前的遗传、分子和功能分析表明,嘈杂基因(cac)在果蝇中编码一种主要的突触前钙通道α1亚基。在此我们报告,与增强型绿色荧光蛋白融合的cac编码的α1亚基的转基因表达有效地挽救了cac致死突变,并允许在体内分析钙通道在活性区的定位。此处报告的结果进一步阐明了cac编码的钙通道在神经递质释放中的主要作用。此外,这些研究为体内功能性突触前钙通道的实时成像提供了一种独特的遗传工具,并定义了一种分子标记,用于相对于活性区对其他突触前蛋白进行免疫定位。预计这些发现将有助于在这个重要的模型系统中对突触发育和功能进行进一步分析。