Microbial light-activatable proton pumps as neuronal inhibitors to functionally dissect neuronal networks in C. elegans.

Essentially any behavior in simple and complex animals depends on neuronal network function. Currently, the best-defined system to study neuronal circuits is the nematode Caenorhabditis elegans, as the connectivity of its 302 neurons is exactly known. Individual neurons can be activated by photostim...

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Autores principales:	Steven J Husson, Jana F Liewald, Christian Schultheis, Jeffrey N Stirman, Hang Lu, Alexander Gottschalk
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2012
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/f08e4009a5b3443088f8b601ae8c934c
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spelling	oai:doaj.org-article:f08e4009a5b3443088f8b601ae8c934c2021-11-18T07:12:21ZMicrobial light-activatable proton pumps as neuronal inhibitors to functionally dissect neuronal networks in C. elegans.1932-620310.1371/journal.pone.0040937https://doaj.org/article/f08e4009a5b3443088f8b601ae8c934c2012-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/22815873/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203Essentially any behavior in simple and complex animals depends on neuronal network function. Currently, the best-defined system to study neuronal circuits is the nematode Caenorhabditis elegans, as the connectivity of its 302 neurons is exactly known. Individual neurons can be activated by photostimulation of Channelrhodopsin-2 (ChR2) using blue light, allowing to directly probe the importance of a particular neuron for the respective behavioral output of the network under study. In analogy, other excitable cells can be inhibited by expressing Halorhodopsin from Natronomonas pharaonis (NpHR) and subsequent illumination with yellow light. However, inhibiting C. elegans neurons using NpHR is difficult. Recently, proton pumps from various sources were established as valuable alternative hyperpolarizers. Here we show that archaerhodopsin-3 (Arch) from Halorubrum sodomense and a proton pump from the fungus Leptosphaeria maculans (Mac) can be utilized to effectively inhibit excitable cells in C. elegans. Arch is the most powerful hyperpolarizer when illuminated with yellow or green light while the action spectrum of Mac is more blue-shifted, as analyzed by light-evoked behaviors and electrophysiology. This allows these tools to be combined in various ways with ChR2 to analyze different subsets of neurons within a circuit. We exemplify this by means of the polymodal aversive sensory ASH neurons, and the downstream command interneurons to which ASH neurons signal to trigger a reversal followed by a directional turn. Photostimulating ASH and subsequently inhibiting command interneurons using two-color illumination of different body segments, allows investigating temporal aspects of signaling downstream of ASH.Steven J HussonJana F LiewaldChristian SchultheisJeffrey N StirmanHang LuAlexander GottschalkPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 7, Iss 7, p e40937 (2012)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Steven J Husson Jana F Liewald Christian Schultheis Jeffrey N Stirman Hang Lu Alexander Gottschalk Microbial light-activatable proton pumps as neuronal inhibitors to functionally dissect neuronal networks in C. elegans.
description	Essentially any behavior in simple and complex animals depends on neuronal network function. Currently, the best-defined system to study neuronal circuits is the nematode Caenorhabditis elegans, as the connectivity of its 302 neurons is exactly known. Individual neurons can be activated by photostimulation of Channelrhodopsin-2 (ChR2) using blue light, allowing to directly probe the importance of a particular neuron for the respective behavioral output of the network under study. In analogy, other excitable cells can be inhibited by expressing Halorhodopsin from Natronomonas pharaonis (NpHR) and subsequent illumination with yellow light. However, inhibiting C. elegans neurons using NpHR is difficult. Recently, proton pumps from various sources were established as valuable alternative hyperpolarizers. Here we show that archaerhodopsin-3 (Arch) from Halorubrum sodomense and a proton pump from the fungus Leptosphaeria maculans (Mac) can be utilized to effectively inhibit excitable cells in C. elegans. Arch is the most powerful hyperpolarizer when illuminated with yellow or green light while the action spectrum of Mac is more blue-shifted, as analyzed by light-evoked behaviors and electrophysiology. This allows these tools to be combined in various ways with ChR2 to analyze different subsets of neurons within a circuit. We exemplify this by means of the polymodal aversive sensory ASH neurons, and the downstream command interneurons to which ASH neurons signal to trigger a reversal followed by a directional turn. Photostimulating ASH and subsequently inhibiting command interneurons using two-color illumination of different body segments, allows investigating temporal aspects of signaling downstream of ASH.
format	article
author	Steven J Husson Jana F Liewald Christian Schultheis Jeffrey N Stirman Hang Lu Alexander Gottschalk
author_facet	Steven J Husson Jana F Liewald Christian Schultheis Jeffrey N Stirman Hang Lu Alexander Gottschalk
author_sort	Steven J Husson
title	Microbial light-activatable proton pumps as neuronal inhibitors to functionally dissect neuronal networks in C. elegans.
title_short	Microbial light-activatable proton pumps as neuronal inhibitors to functionally dissect neuronal networks in C. elegans.
title_full	Microbial light-activatable proton pumps as neuronal inhibitors to functionally dissect neuronal networks in C. elegans.
title_fullStr	Microbial light-activatable proton pumps as neuronal inhibitors to functionally dissect neuronal networks in C. elegans.
title_full_unstemmed	Microbial light-activatable proton pumps as neuronal inhibitors to functionally dissect neuronal networks in C. elegans.
title_sort	microbial light-activatable proton pumps as neuronal inhibitors to functionally dissect neuronal networks in c. elegans.
publisher	Public Library of Science (PLoS)
publishDate	2012
url	https://doaj.org/article/f08e4009a5b3443088f8b601ae8c934c
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Microbial light-activatable proton pumps as neuronal inhibitors to functionally dissect neuronal networks in C. elegans.

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