Zhang, ChuanqiangYang, ShangFlossmann, TomGao, ShiqiangWitte, Otto W.Nagel, GeorgHolthoff, KnutKirmse, Knut2019-12-122019-12-122019-12-122019-11-2710.1186/s12915-019-0717-6https://infoscience.epfl.ch/handle/20.500.14299/163947WOS:000499960800002Background Optogenetic silencing techniques have expanded the causal understanding of the functions of diverse neuronal cell types in both the healthy and diseased brain. A widely used inhibitory optogenetic actuator is eNpHR3.0, an improved version of the light-driven chloride pump halorhodopsin derived from Natronomonas pharaonis. A major drawback of eNpHR3.0 is related to its pronounced inactivation on a time-scale of seconds, which renders it unsuited for applications that require long-lasting silencing. Results Using transgenic mice and Xenopus laevis oocytes expressing an eNpHR3.0-EYFP fusion protein, we here report optimized photo-stimulation techniques that profoundly increase the stability of eNpHR3.0-mediated currents during long-term photo-stimulation. We demonstrate that optimized photo-stimulation enables prolonged hyperpolarization and suppression of action potential discharge on a time-scale of minutes. Conclusions Collectively, our findings extend the utility of eNpHR3.0 to the long-lasting inhibition of excitable cells, thus facilitating the optogenetic dissection of neural circuits.BiologyLife Sciences & Biomedicine - Other Topicshalorhodopsinenphr30optogeneticinhibitiontransgeniclight-driven protoncrystal-structuresgabaergic neuronschloridepharaonisbacteriorhodopsindeprotonationinterneuronsactivationdependencetext::journal::journal article::research article