Institute of Biophysical Chemistry and Buchmann Institute for Molecular Life Sciences, Johann Wolfgang Goethe-University, Frankfurt, Germany.
Methods Mol Biol. 2022;2483:61-76. doi: 10.1007/978-1-0716-2245-2_4.
In the past 15 years, optogenetic methods became invaluable tools in neurobiological research but also in general cell biology. Most prominently, optogenetic methods utilize microbial rhodopsins to elicit neuronal de- or hyperpolarization. However, other optogenetic tools have emerged that allow influencing neuronal function by different approaches. In this chapter we describe the use of photoactivated adenylyl cyclases (PACs) as modulators of neuronal activity. Using Caenorhabditis elegans as a model organism, this chapter shows how to measure the effect of PAC photoactivation by behavioral assays in different tissues (neurons and muscles), as well as their significance to neurobiology. Further, this chapter describes in vitro cyclic nucleoside-3',5'-monophosphate measurements (cNMP) to characterize new PACs in C. elegans.
在过去的 15 年中,光遗传学方法已成为神经生物学研究以及一般细胞生物学领域中不可或缺的工具。其中最为突出的是,光遗传学方法利用微生物视紫红质来引发神经元去极化或超极化。然而,其他光遗传学工具的出现也使得通过不同的方法来影响神经元功能成为可能。在本章中,我们将描述光激活腺苷酸环化酶(PACs)作为神经元活动调节剂的应用。我们使用秀丽隐杆线虫作为模式生物,展示了如何通过行为测定法在不同组织(神经元和肌肉)中测量 PAC 光激活的效果,以及它们对神经生物学的意义。此外,本章还描述了体外环核苷酸-3',5'-单磷酸测量(cNMP),以鉴定秀丽隐杆线虫中的新型 PAC。