Forward and reverse trafficking of L-type calcium channels

钙离子对生理功能的各个方面都很重要，包括记忆、神经信号的传输和心跳的产生。钙进入细胞是由钙通道介导的，钙通道是嵌入细胞膜的一系列蛋白质，允许钙离子通过细胞膜。由于过高的钙水平可能对细胞有害，因此通过这些渠道进入的钙量受到严格控制。因此，考虑到它们的关键作用，了解钙通道如何在分子水平上发挥作用，以及细胞如何调节存在的钙通道蛋白的量是至关重要的。在各种类型的钙通道中，一类特别重要的是L型钙通道，它对基因和钙依赖酶的激活至关重要。在这一应用中，我们建议研究控制L类钙通道从合成和组装的细胞内转运到调节钙内流的细胞膜的分子基础。我们还将确定细胞活动如何调节通道在细胞表面停留的时间。我们将研究调节蛋白和辅助钙通道亚单位是如何影响这种运输的。我们将结合使用最先进的实验技术来实现我们的目标。这些包括分子生物学、生物化学、光遗传学和模型系统和神经元中钙通道的电子记录。具体地说，我们将通过DNA技术操纵钙通道及其调节蛋白的功能，然后研究这如何影响它们的组装、运输和膜的稳定性。总而言之，这项工作将为一个适用于广泛物种的基本生物过程提供新的见解。

Calcium ions are important for a variety of aspects of physiological function including memory, transmission of nerve signals, and the generation of the heart beat. Calcium entry into cells is mediated by calcium channels, a family of proteins which are embedded in the cell membrane and allow calcium ions to pass through them. Because excessive calcium levels can be toxic to cells, the amount of calcium entering through these channels is tightly regulated. Given their pivotal roles, it is therefore critical to understand how the calcium channels function at the molecular level, and how cells regulate the amount of calcium channel protein that is present. Among the various types of calcium channels, one particularly important class are the L-type calcium channels which are vital for the activation of genes and calcium dependent enzymes. In this application, we propose to investigate the molecular basis that governs the trafficking of L-type calcium channels from the inside of the cells where they are synthesized and assembled, to the cell membrane where they regulate calcium entry. We also will determine how cellular activity regulates how long the channels remain at the cell surface. We will investigate how regulatory proteins and auxiliary calcium channels subunits that form part of the channel affect this trafficking. We will use a combination of state of the art experimental techniques to tackle our objectives. These include molecular biology, biochemistry, optogenetics and electrical recordings from calcium channels in model systems and in neurons. Specifically, we will manipulate the function of calcium channels and their regulatory proteins via DNA technology, and then examine how this affects their assembly, trafficking, and membrane stability. Altogether, this work will provide novel insights about a fundamental biological process that is applicable across a wide range of species.