title: Optogenetic inhibition of neurons by internal light production.
abstract: Optogenetics is an extremely powerful tool for selective neuronal activation/inhibition and dissection of neural circuits. However, a limitation of in vivo optogenetics is that an animal must be tethered to an optical fiber for delivery of light. Here, we describe a new method for in vivo, optogenetic inhibition of neural activity using an internal, animal-generated light source based on firefly luciferase. Two adeno-associated viruses encoding luciferase were tested and both produced concentration-dependent light after administration of the substrate, luciferin. Mice were co-infected with halorhodopsin- and luciferase-expressing viruses in the striatum, and luciferin administration significantly reduced Fos activity compared to control animals infected with halorhodopsin only. Recordings of neuronal activity in behaving animals confirmed that firing was greatly reduced after luciferin administration. Finally, amphetamine-induced locomotor activity was reduced in halorhodopsin/luciferase mice pre-injected with luciferin compared to controls. This demonstrates that virally encoded luciferase is able to generate sufficient light to activate halorhodopsin and suppress neural activity and change behavior. This approach could be used to generate inhibition in response to activation of specific molecular pathways.
authors.affiliation: Department of Psychiatry, Yale University School of Medicine New Haven, CT, USA.
authors.forename: Benjamin B
authors.initials: BB
authors.lastname: Land
authors.affiliation: Department of Psychiatry, Yale University School of Medicine New Haven, CT, USA.
authors.forename: Catherine E
authors.initials: CE
authors.lastname: Brayton
authors.affiliation: Department of Psychiatry, Yale University School of Medicine New Haven, CT, USA.
authors.forename: Kara E
authors.initials: KE
authors.lastname: Furman
authors.affiliation: Department of Psychiatry, Yale University School of Medicine New Haven, CT, USA.
authors.forename: Zoe
authors.initials: Z
authors.lastname: Lapalombara
authors.affiliation: Department of Psychiatry, Yale University School of Medicine New Haven, CT, USA.
authors.forename: Ralph J
authors.initials: RJ
authors.lastname: Dileone
keywords: amphetamine; dopamine; luciferase; optogenetics; striatum
references.citation: Annu Rev Pathol. 2007;2:277-305
references.pmid: 18039101
references.citation: J Neurosci. 2010 Jun 16;30(24):8263-73
references.pmid: 20554878
references.citation: Neuron. 2009 Jul 16;63(1):27-39
references.pmid: 19607790
references.citation: Proc Natl Acad Sci U S A. 2007 Mar 20;104(12):5163-8
references.pmid: 17360345
references.citation: Science. 2013 Apr 12;340(6129):211-6
references.pmid: 23580530
references.citation: Nature. 2013 Nov 7;503(7474):111-4
references.pmid: 24121436
references.citation: Nature. 2007 Apr 5;446(7136):633-9
references.pmid: 17410168
references.citation: Nat Med. 2003 Dec;9(12):1539-44
references.pmid: 14634645
references.citation: Nature. 2006 Mar 16;440(7082):372-6
references.pmid: 16541080
references.citation: PLoS One. 2011;6(5):e20360
references.pmid: 21647372
references.citation: Science. 2010 Dec 17;330(6011):1677-81
references.pmid: 21164015
references.citation: Proc Natl Acad Sci U S A. 2012 Dec 11;109(50):20726-31
references.pmid: 23185016
references.citation: J Neural Eng. 2011 Aug;8(4):046021
references.pmid: 21701058
references.citation: Methods Enzymol. 2012;506:117-33
references.pmid: 22341222
references.citation: Biotechniques. 2003 Jun;34(6):1184-8
references.pmid: 12813886
references.citation: Science. 2012 Mar 23;335(6075):1513-6
references.pmid: 22442487