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    Lehrstuhl für Physiologie - Schwerpunkt Neurophysiologie

    Optimized photo-stimulation of halorhodopsin for long-term neuronal inhibition

    27.11.2019

    Optimized photo-stimulation of halorhodopsin for long-term neuronal inhibition

     

    Chuanqiang Zhang, Shang Yang, Tom Flossmann, Shiqiang Gao, Otto W. Witte, Georg Nagel, Knut Holthoff, Knut Kirmse (2019)

    BMC Biology volume 17, 95 

    Background: 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.

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