Diminishing neuronal acidification by channelrhodopsins with low proton conduction

Author:

Hayward Rebecca Frank1ORCID,Brooks F Phil2,Yang Shang3,Gao Shiqiang3ORCID,Cohen Adam E24ORCID

Affiliation:

1. School of Engineering and Applied Sciences, Harvard University

2. Department of Chemistry, Harvard University

3. Department of Neurophysiology, University of Wurzburg

4. Department of Physics, Harvard University

Abstract

Many channelrhodopsins are permeable to protons. We found that in neurons, activation of a high-current channelrhodopsin, CheRiff, led to significant acidification, with faster acidification in the dendrites than in the soma. Experiments with patterned optogenetic stimulation in monolayers of HEK cells established that the acidification was due to proton transport through the opsin, rather than through other voltage-dependent channels. We identified and characterized two opsins which showed large photocurrents, but small proton permeability, PsCatCh2.0 and ChR2-3M. PsCatCh2.0 showed excellent response kinetics and was also spectrally compatible with simultaneous voltage imaging with QuasAr6a. Stimulation-evoked acidification is a possible source of disruptions to cell health in scientific and prospective therapeutic applications of optogenetics. Channelrhodopsins with low proton permeability are a promising strategy for avoiding these problems.

Funder

Office of Naval Research

National Institutes of Health

Publisher

eLife Sciences Publications, Ltd

Subject

General Immunology and Microbiology,General Biochemistry, Genetics and Molecular Biology,General Medicine,General Neuroscience

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