A light-gated potassium channel for sustained neuronal inhibition

Laura Alberio; Andrea Locarno; Andrea Saponaro; Edoardo Romano; Valérie Bercier; Shahad Albadri; Federica Simeoni; Silvia Moleri; Silvia Pelucchi; Alessandro Porro; Elena Marcello; Noemi Barsotti; Kerri Kukovetz; Arjen J. Boender; Andrea Contestabile; Shizhen Luo; Aubin Moutal; Yingshi Ji; Giulia Romani; Monica Beltrame; Filippo Del Bene; Monica Di Luca; Rajesh Khanna; Henry M. Colecraft; Massimo Pasqualetti; Gerhard Thiel; Raffaella Tonini; Anna Moroni

doi:10.1038/s41592-018-0186-9

A light-gated potassium channel for sustained neuronal inhibition

Laura Alberio, Andrea Locarno, Andrea Saponaro, Edoardo Romano, Valérie Bercier, Shahad Albadri, Federica Simeoni, Silvia Moleri, Silvia Pelucchi, Alessandro Porro, Elena Marcello, Noemi Barsotti, Kerri Kukovetz, Arjen J. Boender, Andrea Contestabile, Shizhen Luo, Aubin Moutal, Yingshi Ji, Giulia Romani, Monica BeltrameFilippo Del Bene, Monica Di Luca, Rajesh Khanna, Henry M. Colecraft, Massimo Pasqualetti, Gerhard Thiel, Raffaella Tonini, Anna Moroni

Molecular Pathobiology

Research output: Contribution to journal › Article › peer-review

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.

Original language	English (US)
Pages (from-to)	969-976
Number of pages	8
Journal	Nature methods
Volume	15
Issue number	11
DOIs	https://doi.org/10.1038/s41592-018-0186-9
State	Published - Nov 1 2018

ASJC Scopus subject areas

Biotechnology
Biochemistry
Molecular Biology
Cell Biology

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10.1038/s41592-018-0186-9

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Alberio, L., Locarno, A., Saponaro, A., Romano, E., Bercier, V., Albadri, S., Simeoni, F., Moleri, S., Pelucchi, S., Porro, A., Marcello, E., Barsotti, N., Kukovetz, K., Boender, A. J., Contestabile, A., Luo, S., Moutal, A., Ji, Y., Romani, G., ... Moroni, A. (2018). A light-gated potassium channel for sustained neuronal inhibition. Nature methods, 15(11), 969-976. https://doi.org/10.1038/s41592-018-0186-9

Alberio, L, Locarno, A, Saponaro, A, Romano, E, Bercier, V, Albadri, S, Simeoni, F, Moleri, S, Pelucchi, S, Porro, A, Marcello, E, Barsotti, N, Kukovetz, K, Boender, AJ, Contestabile, A, Luo, S, Moutal, A, Ji, Y, Romani, G, Beltrame, M, Del Bene, F, Di Luca, M, Khanna, R, Colecraft, HM, Pasqualetti, M, Thiel, G, Tonini, R & Moroni, A 2018, 'A light-gated potassium channel for sustained neuronal inhibition', Nature methods, vol. 15, no. 11, pp. 969-976. https://doi.org/10.1038/s41592-018-0186-9

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title = "A light-gated potassium channel for sustained neuronal inhibition",

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.",

author = "Laura Alberio and Andrea Locarno and Andrea Saponaro and Edoardo Romano and Val{\'e}rie Bercier and Shahad Albadri and Federica Simeoni and Silvia Moleri and Silvia Pelucchi and Alessandro Porro and Elena Marcello and Noemi Barsotti and Kerri Kukovetz and Boender, {Arjen J.} and Andrea Contestabile and Shizhen Luo and Aubin Moutal and Yingshi Ji and Giulia Romani and Monica Beltrame and {Del Bene}, Filippo and {Di Luca}, Monica and Rajesh Khanna and Colecraft, {Henry M.} and Massimo Pasqualetti and Gerhard Thiel and Raffaella Tonini and Anna Moroni",

note = "Funding Information: We thank S. Guazzi and M. Festa for technical help with cloning and zebrafish expression. We acknowledge M. Pesce and A. Gino for help with immunohistochemistry. pENN-AAV-hSyn-Cre-WPRE-hGH was a gift from J.M. Wilson (Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA). This work was supported by the 2016 Schaefer Research Scholars Program of Columbia University (to A. Moroni), MIUR PRIN (Programmi di Ricerca di Rilevante Interesse Nazionale; 494 2015, 2015795S5W to A. Moroni), the European Research Council (ERC; 2015 Advanced Grant 495 (AdG) n. 695078 noMAGIC to A. Moroni and G.T.), DFG priority program SPP1926 (to G.T.), the Fondazione Istituto Italiano di Tecnologia (to A.L., A.C., A.J.B. and R.T.), and AIRAlzh Onlus-COOP Italia (fellowship to S.P.). Publisher Copyright: {\textcopyright} 2018, The Author(s), under exclusive licence to Springer Nature America, Inc.",

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T1 - A light-gated potassium channel for sustained neuronal inhibition

AU - Alberio, Laura

AU - Locarno, Andrea

AU - Saponaro, Andrea

AU - Romano, Edoardo

AU - Bercier, Valérie

AU - Albadri, Shahad

AU - Simeoni, Federica

AU - Moleri, Silvia

AU - Pelucchi, Silvia

AU - Porro, Alessandro

AU - Marcello, Elena

AU - Barsotti, Noemi

AU - Kukovetz, Kerri

AU - Boender, Arjen J.

AU - Contestabile, Andrea

AU - Luo, Shizhen

AU - Moutal, Aubin

AU - Ji, Yingshi

AU - Romani, Giulia

AU - Beltrame, Monica

AU - Del Bene, Filippo

AU - Di Luca, Monica

AU - Khanna, Rajesh

AU - Colecraft, Henry M.

AU - Pasqualetti, Massimo

AU - Thiel, Gerhard

AU - Tonini, Raffaella

AU - Moroni, Anna

N1 - Funding Information: We thank S. Guazzi and M. Festa for technical help with cloning and zebrafish expression. We acknowledge M. Pesce and A. Gino for help with immunohistochemistry. pENN-AAV-hSyn-Cre-WPRE-hGH was a gift from J.M. Wilson (Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA). This work was supported by the 2016 Schaefer Research Scholars Program of Columbia University (to A. Moroni), MIUR PRIN (Programmi di Ricerca di Rilevante Interesse Nazionale; 494 2015, 2015795S5W to A. Moroni), the European Research Council (ERC; 2015 Advanced Grant 495 (AdG) n. 695078 noMAGIC to A. Moroni and G.T.), DFG priority program SPP1926 (to G.T.), the Fondazione Istituto Italiano di Tecnologia (to A.L., A.C., A.J.B. and R.T.), and AIRAlzh Onlus-COOP Italia (fellowship to S.P.). Publisher Copyright: © 2018, The Author(s), under exclusive licence to Springer Nature America, Inc.

PY - 2018/11/1

Y1 - 2018/11/1

N2 - 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.

AB - 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.

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DO - 10.1038/s41592-018-0186-9

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AN - SCOPUS:85055765457

SN - 1548-7091

VL - 15

SP - 969

EP - 976

JO - Nature methods

JF - Nature methods

IS - 11

ER -

A light-gated potassium channel for sustained neuronal inhibition

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