Optimized photo-stimulation of halorhodopsin for long-term neuronal inhibition.
Halorhodopsin
Inhibition
Optogenetic
Transgenic
eNpHR3.0
Journal
BMC biology
ISSN: 1741-7007
Titre abrégé: BMC Biol
Pays: England
ID NLM: 101190720
Informations de publication
Date de publication:
27 11 2019
27 11 2019
Historique:
received:
30
09
2019
accepted:
30
10
2019
entrez:
29
11
2019
pubmed:
30
11
2019
medline:
12
5
2020
Statut:
epublish
Résumé
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. 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. 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.
Sections du résumé
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.
Identifiants
pubmed: 31775747
doi: 10.1186/s12915-019-0717-6
pii: 10.1186/s12915-019-0717-6
pmc: PMC6882325
doi:
Substances chimiques
Bacterial Proteins
0
Halorhodopsins
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
95Références
Front Neurosci. 2018 Oct 02;12:643
pubmed: 30333716
PLoS One. 2007 Mar 21;2(3):e299
pubmed: 17375185
Proc Natl Acad Sci U S A. 2016 Jan 26;113(4):822-9
pubmed: 26699459
Proc Natl Acad Sci U S A. 1993 Jan 15;90(2):639-43
pubmed: 8380643
EMBO J. 1983;2(8):1379-85
pubmed: 10872334
Neuron. 2014 Mar 19;81(6):1263-1273
pubmed: 24656249
Neuron. 2017 Aug 2;95(3):504-529
pubmed: 28772120
Science. 2006 Jan 13;311(5758):233-5
pubmed: 16410524
Science. 2015 Aug 7;349(6248):647-50
pubmed: 26113638
Nature. 2009 Apr 23;458(7241):1025-9
pubmed: 19295515
Proc Natl Acad Sci U S A. 2009 Jul 21;106(29):12162-7
pubmed: 19581573
J Neurosci. 2015 May 20;35(20):7715-26
pubmed: 25995461
Proc Natl Acad Sci U S A. 2017 Dec 12;114(50):E10819-E10828
pubmed: 29183979
J Mol Biol. 2010 Feb 26;396(3):564-79
pubmed: 19961859
Proc Natl Acad Sci U S A. 2009 Oct 20;106(42):17968-73
pubmed: 19805086
Biochemistry. 2005 Feb 22;44(7):2284-92
pubmed: 15709741
J Physiol. 1989 Sep;416:303-25
pubmed: 2575165
Nat Commun. 2018 Nov 5;9(1):4611
pubmed: 30397200
Nat Neurosci. 2016 Apr;19(4):554-6
pubmed: 26950004
J Neurosci. 2006 Nov 1;26(44):11432-6
pubmed: 17079672
Nat Med. 2005 Nov;11(11):1205-13
pubmed: 16227993
Cell. 2010 Apr 2;141(1):154-165
pubmed: 20303157
Biophys J. 2001 Sep;81(3):1600-12
pubmed: 11509373
J Mol Biol. 2011 Oct 14;413(1):162-76
pubmed: 21871461
Nat Neurosci. 2014 Aug;17(8):1123-9
pubmed: 24997763
Neuron. 2011 Sep 22;71(6):995-1013
pubmed: 21943598
Nat Neurosci. 2019 Jul;22(7):1061-1065
pubmed: 31209378
Cell. 2011 Dec 23;147(7):1446-57
pubmed: 22196724
Nat Neurosci. 2012 Jun 24;15(8):1102-4
pubmed: 22729174
Nat Methods. 2019 Nov;16(11):1176-1184
pubmed: 31611694
Nature. 2010 Jan 7;463(7277):98-102
pubmed: 20054397
Biochemistry. 2005 Nov 1;44(43):14231-7
pubmed: 16245939
Cell Calcium. 2012 Aug;52(2):182-9
pubmed: 22658827
Curr Opin Neurobiol. 2015 Oct;34:29-36
pubmed: 25637880
Biochemistry. 1986 Oct 21;25(21):6706-11
pubmed: 3790553
Biophys J. 2015 Jun 2;108(11):2680-90
pubmed: 26039169
Biochemistry. 1995 Nov 7;34(44):14490-9
pubmed: 7578054
Cell Rep. 2019 Mar 19;26(12):3173-3182.e5
pubmed: 30893591
Front Syst Neurosci. 2016 Sep 02;10:74
pubmed: 27642278
Nat Commun. 2015 Sep 28;6:8480
pubmed: 26412387
EMBO J. 1985 Sep;4(9):2347-50
pubmed: 15938053
Epilepsy Res. 2008 May;79(2-3):201-12
pubmed: 18394864
J Neurosci. 2002 Aug 1;22(15):6309-14
pubmed: 12151506
Neuroscientist. 2017 Feb;23(1):68-81
pubmed: 26700888
Nature. 2007 Apr 5;446(7136):633-9
pubmed: 17410168
Eur J Biochem. 1973 Aug 17;37(2):316-26
pubmed: 4745733
Nat Neurosci. 2012 Mar 25;15(5):793-802
pubmed: 22446880
Nat Methods. 2011 Dec 18;9(2):159-72
pubmed: 22179551
FEBS Lett. 1995 Dec 18;377(2):263-6
pubmed: 8543064
Sci Rep. 2015 Oct 07;5:14807
pubmed: 26443033
Biochim Biophys Acta. 1978 Aug 8;503(2):304-15
pubmed: 28756