Optogenetic manipulation of cerebellar Purkinje cell activity in vivo.

Purkinje cells (PCs) are the sole output neurons of the cerebellar cortex. Although their anatomical connections and physiological response properties have been extensively studied, the causal role of their activity in behavioral, cognitive and autonomic functions is still unclear because PC activit...

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Autores principales:	Tadashi Tsubota, Yohei Ohashi, Keita Tamura, Ayana Sato, Yasushi Miyashita
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2011
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/83160ac402994c3186e676def780cc7f
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spelling	oai:doaj.org-article:83160ac402994c3186e676def780cc7f2021-11-18T06:48:35ZOptogenetic manipulation of cerebellar Purkinje cell activity in vivo.1932-620310.1371/journal.pone.0022400https://doaj.org/article/83160ac402994c3186e676def780cc7f2011-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/21850224/?tool=EBIhttps://doaj.org/toc/1932-6203Purkinje cells (PCs) are the sole output neurons of the cerebellar cortex. Although their anatomical connections and physiological response properties have been extensively studied, the causal role of their activity in behavioral, cognitive and autonomic functions is still unclear because PC activity cannot be selectively controlled. Here we developed a novel technique using optogenetics for selective and rapidly reversible manipulation of PC activity in vivo. We injected into rat cerebellar cortex lentiviruses expressing either the light-activated cationic channel channelrhodopsin-2 (ChR2) or light-driven chloride pump halorhodopsin (eNpHR) under the control of the PC-specific L7 promoter. Transgene expression was observed in most PCs (ChR2, 92.6%; eNpHR, 95.3%), as determined by immunohistochemical analysis. In vivo electrophysiological recordings showed that all light-responsive PCs in ChR2-transduced rats increased frequency of simple spike in response to blue laser illumination. Similarly, most light-responsive PCs (93.8%) in eNpHR-transduced rats decreased frequency of simple spike in response to orange laser illumination. We then applied these techniques to characterize the roles of rat cerebellar uvula, one of the cardiovascular regulatory regions in the cerebellum, in resting blood pressure (BP) regulation in anesthetized rats. ChR2-mediated photostimulation and eNpHR-mediated photoinhibition of the uvula had opposite effects on resting BP, inducing depressor and pressor responses, respectively. In contrast, manipulation of PC activity within the neighboring lobule VIII had no effect on BP. Blue and orange laser illumination onto PBS-injected lobule IX didn't affect BP, indicating the observed effects on BP were actually due to PC activation and inhibition. These results clearly demonstrate that the optogenetic method we developed here will provide a powerful way to elucidate a causal relationship between local PC activity and functions of the cerebellum.Tadashi TsubotaYohei OhashiKeita TamuraAyana SatoYasushi MiyashitaPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 6, Iss 8, p e22400 (2011)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Tadashi Tsubota Yohei Ohashi Keita Tamura Ayana Sato Yasushi Miyashita Optogenetic manipulation of cerebellar Purkinje cell activity in vivo.
description	Purkinje cells (PCs) are the sole output neurons of the cerebellar cortex. Although their anatomical connections and physiological response properties have been extensively studied, the causal role of their activity in behavioral, cognitive and autonomic functions is still unclear because PC activity cannot be selectively controlled. Here we developed a novel technique using optogenetics for selective and rapidly reversible manipulation of PC activity in vivo. We injected into rat cerebellar cortex lentiviruses expressing either the light-activated cationic channel channelrhodopsin-2 (ChR2) or light-driven chloride pump halorhodopsin (eNpHR) under the control of the PC-specific L7 promoter. Transgene expression was observed in most PCs (ChR2, 92.6%; eNpHR, 95.3%), as determined by immunohistochemical analysis. In vivo electrophysiological recordings showed that all light-responsive PCs in ChR2-transduced rats increased frequency of simple spike in response to blue laser illumination. Similarly, most light-responsive PCs (93.8%) in eNpHR-transduced rats decreased frequency of simple spike in response to orange laser illumination. We then applied these techniques to characterize the roles of rat cerebellar uvula, one of the cardiovascular regulatory regions in the cerebellum, in resting blood pressure (BP) regulation in anesthetized rats. ChR2-mediated photostimulation and eNpHR-mediated photoinhibition of the uvula had opposite effects on resting BP, inducing depressor and pressor responses, respectively. In contrast, manipulation of PC activity within the neighboring lobule VIII had no effect on BP. Blue and orange laser illumination onto PBS-injected lobule IX didn't affect BP, indicating the observed effects on BP were actually due to PC activation and inhibition. These results clearly demonstrate that the optogenetic method we developed here will provide a powerful way to elucidate a causal relationship between local PC activity and functions of the cerebellum.
format	article
author	Tadashi Tsubota Yohei Ohashi Keita Tamura Ayana Sato Yasushi Miyashita
author_facet	Tadashi Tsubota Yohei Ohashi Keita Tamura Ayana Sato Yasushi Miyashita
author_sort	Tadashi Tsubota
title	Optogenetic manipulation of cerebellar Purkinje cell activity in vivo.
title_short	Optogenetic manipulation of cerebellar Purkinje cell activity in vivo.
title_full	Optogenetic manipulation of cerebellar Purkinje cell activity in vivo.
title_fullStr	Optogenetic manipulation of cerebellar Purkinje cell activity in vivo.
title_full_unstemmed	Optogenetic manipulation of cerebellar Purkinje cell activity in vivo.
title_sort	optogenetic manipulation of cerebellar purkinje cell activity in vivo.
publisher	Public Library of Science (PLoS)
publishDate	2011
url	https://doaj.org/article/83160ac402994c3186e676def780cc7f
work_keys_str_mv	AT tadashitsubota optogeneticmanipulationofcerebellarpurkinjecellactivityinvivo AT yoheiohashi optogeneticmanipulationofcerebellarpurkinjecellactivityinvivo AT keitatamura optogeneticmanipulationofcerebellarpurkinjecellactivityinvivo AT ayanasato optogeneticmanipulationofcerebellarpurkinjecellactivityinvivo AT yasushimiyashita optogeneticmanipulationofcerebellarpurkinjecellactivityinvivo
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Optogenetic manipulation of cerebellar Purkinje cell activity in vivo.

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