Author:

Keywords:

Science & Technology, Life Sciences & Biomedicine, Biochemical Research Methods, Biochemistry & Molecular Biology, 14-3-3 PROTEINS, IN-VIVO, PERIPHERAL NEUROPATHY, CHLORIDE CHANNEL, NEURAL CIRCUITRY, BRAIN-SLICES, CHANNELRHODOPSIN, KAT1, TRANSMISSION, CONSTRAINTS, Action Potentials, Animals, Female, Hyperalgesia, Light, Male, Mice, Inbred C57BL, Neurons, Optogenetics, Paclitaxel, Pain, Peripheral Nervous System Diseases, Rats, Rats, Sprague-Dawley, Recombinant Fusion Proteins, Zebrafish, 06 Biological Sciences, 10 Technology, 11 Medical and Health Sciences, Developmental Biology, 31 Biological sciences

Abstract:

Currently available inhibitory optogenetic tools provide short and transient silencing of neurons, but they cannot provide long-lasting inhibition because of the requirement for high light intensities. Here we present an optimized blue-light-sensitive synthetic potassium channel, BLINK2, which showed good expression in neurons in three species. The channel is activated by illumination with low doses of blue light, and in our experiments it remained active over (tens of) minutes in the dark after the illumination was stopped. This activation caused long periods of inhibition of neuronal firing in ex vivo recordings of mouse neurons and impaired motor neuron response in zebrafish in vivo. As a proof-of-concept application, we demonstrated that in a freely moving rat model of neuropathic pain, the activation of a small number of BLINK2 channels caused a long-lasting (>30 min) reduction in pain sensation.