Reinbothe Thomas M, Mollet Inês G
Department of Physiology, University of Gothenburg, Box 432, Medicinaregatan 11-13, 40530, Gothenburg, Sweden.
Department of Clinical Sciences, Malmö, Lund University Diabetes Centre, Lund University, Malmö, Sweden.
Methods Mol Biol. 2016;1408:107-23. doi: 10.1007/978-1-4939-3512-3_8.
In light of the emerging diabetes epidemic, new experimental approaches in islet research are needed to elucidate the mechanisms behind pancreatic islet dysfunction and to facilitate the development of more effective therapies. Optogenetics has created numerous new experimental tools enabling us to gain insights into processes little was known about before. The spatial and temporal precision that it can achieve is also attractive for studying the cells of the pancreatic islet and we set out to explore the possibilities of this technology for our purposes. We here describe how to use the islets of an "optogenetic beta-cell" mouse line in islet batch incubations and Ca(2+) imaging experiments. This protocol enables light-induced insulin release and provides an all-optical solution to control and measure intracellular Ca(2+) levels in pancreatic beta-cells. The technique is easy to set up and provides a useful tool for controlling the activity of distinct islet cell populations.
鉴于糖尿病的流行趋势不断加剧，胰岛研究需要新的实验方法来阐明胰岛功能障碍背后的机制，并促进更有效治疗方法的开发。光遗传学创造了许多新的实验工具，使我们能够深入了解以前知之甚少的过程。它所能实现的时空精度对于研究胰岛细胞也很有吸引力，因此我们着手探索这项技术用于我们研究目的的可能性。我们在此描述如何在胰岛批量培养和Ca(2+)成像实验中使用“光遗传学β细胞”小鼠系的胰岛。该方案能够实现光诱导的胰岛素释放，并提供一种全光学解决方案来控制和测量胰腺β细胞内的Ca(2+)水平。该技术易于设置，为控制不同胰岛细胞群的活性提供了一个有用的工具。
