Show simple item record

2021-05-06Zeitschriftenartikel DOI: 10.1111/bph.15445
Optogenetic tools for manipulation of cyclic nucleotides functionally coupled to cyclic nucleotide‐gated channels
dc.contributor.authorHenß, Thilo
dc.contributor.authorNagpal, Jatin
dc.contributor.authorGao, Shiqiang
dc.contributor.authorScheib, Ulrike
dc.contributor.authorPieragnolo, Alessia
dc.contributor.authorHirschhäuser, Alexander
dc.contributor.authorSchneider-Warme, Franziska
dc.contributor.authorHegemann, Peter
dc.contributor.authorNagel, Georg
dc.contributor.authorGottschalk, Alexander
dc.date.accessioned2021-10-27T08:10:44Z
dc.date.available2021-10-27T08:10:44Z
dc.date.issued2021-05-06none
dc.date.updated2021-06-01T11:59:55Z
dc.identifier.urihttp://edoc.hu-berlin.de/18452/24268
dc.description.abstractBackground and Purpose The cyclic nucleotides cAMP and cGMP are ubiquitous second messengers regulating numerous biological processes. Malfunctional cNMP signalling is linked to diseases and thus is an important target in pharmaceutical research. The existing optogenetic toolbox in Caenorhabditis elegans is restricted to soluble adenylyl cyclases, the membrane‐bound Blastocladiella emersonii CyclOp and hyperpolarizing rhodopsins; yet missing are membrane‐bound photoactivatable adenylyl cyclases and hyperpolarizers based on K+ currents. Experimental Approach For the characterization of photoactivatable nucleotidyl cyclases, we expressed the proteins alone or in combination with cyclic nucleotide‐gated channels in muscle cells and cholinergic motor neurons. To investigate the extent of optogenetic cNMP production and the ability of the systems to depolarize or hyperpolarize cells, we performed behavioural analyses, measured cNMP content in vitro, and compared in vivo expression levels. Key Results We implemented Catenaria CyclOp as a new tool for cGMP production, allowing fine‐control of cGMP levels. We established photoactivatable membrane‐bound adenylyl cyclases, based on mutated versions (“A‐2x”) of Blastocladiella and Catenaria (“Be,” “Ca”) CyclOp, as N‐terminal YFP fusions, enabling more efficient and specific cAMP signalling compared to soluble bPAC, despite lower overall cAMP production. For hyperpolarization of excitable cells by two‐component optogenetics, we introduced the cAMP‐gated K+‐channel SthK from Spirochaeta thermophila and combined it with bPAC, BeCyclOp(A‐2x), or YFP‐BeCyclOp(A‐2x). As an alternative, we implemented the B. emersonii cGMP‐gated K+‐channel BeCNG1 together with BeCyclOp. Conclusion and Implications We established a comprehensive suite of optogenetic tools for cNMP manipulation, applicable in many cell types, including sensory neurons, and for potent hyperpolarization.eng
dc.description.sponsorshipDeutsche Forschungsgemeinschaft (DFG) http://dx.doi.org/10.13039/501100001659
dc.language.isoengnone
dc.publisherHumboldt-Universität zu Berlin
dc.rights(CC BY-NC-ND 4.0) Attribution-NonCommercial-NoDerivatives 4.0 Internationalger
dc.rights.urihttps://creativecommons.org/licenses/by-nc-nd/4.0/
dc.subjectadenylyl cyclaseseng
dc.subjectCaenorhabditis eleganseng
dc.subjectcyclic nucleotide‐gated channelseng
dc.subjectguanylyl cyclaseseng
dc.subjectneuromuscular systemeng
dc.subjectoptogeneticseng
dc.subjectrhodopsineng
dc.subject.ddc610 Medizin und Gesundheitnone
dc.titleOptogenetic tools for manipulation of cyclic nucleotides functionally coupled to cyclic nucleotide‐gated channelsnone
dc.typearticle
dc.identifier.urnurn:nbn:de:kobv:11-110-18452/24268-5
dc.identifier.doi10.1111/bph.15445none
dc.identifier.doihttp://dx.doi.org/10.18452/23610
dc.type.versionpublishedVersionnone
local.edoc.container-titleBritish Journal of Pharmacologynone
local.edoc.pages19none
local.edoc.type-nameZeitschriftenartikel
local.edoc.institutionMathematisch-Naturwissenschaftliche Fakultätnone
local.edoc.container-typeperiodical
local.edoc.container-type-nameZeitschrift
local.edoc.container-publisher-nameWileynone
local.edoc.container-publisher-placeMaldennone
local.edoc.container-firstpage1none
local.edoc.container-lastpage19none
dc.description.versionPeer Reviewednone
local.edoc.affiliationHenß, Thilo; Buchmann Institute for Molecular Life Sciences Goethe University Frankfurt Germanynone
local.edoc.affiliationNagpal, Jatin; Buchmann Institute for Molecular Life Sciences Goethe University Frankfurt Germanynone
local.edoc.affiliationGao, Shiqiang; Department of Neurophysiology, Institute of Physiology, Biocentre Julius‐Maximilians‐University Würzburg Germanynone
local.edoc.affiliationScheib, Ulrike; Institute for Biology, Experimental Biophysics Humboldt‐Universität zu Berlin Berlin Germanynone
local.edoc.affiliationPieragnolo, Alessia; Faculty of Pharmacy University of Padova Padova Italynone
local.edoc.affiliationHirschhäuser, Alexander; Buchmann Institute for Molecular Life Sciences Goethe University Frankfurt Germanynone
local.edoc.affiliationSchneider‐Warme, Franziska; University Heart Center, Medical Center – University of Freiburg and Faculty of Medicine Institute for Experimental Cardiovascular Medicine Freiburg Germanynone
local.edoc.affiliationHegemann, Peter; Institute for Biology, Experimental Biophysics Humboldt‐Universität zu Berlin Berlin Germanynone
local.edoc.affiliationNagel, Georg; Department of Neurophysiology, Institute of Physiology, Biocentre Julius‐Maximilians‐University Würzburg Germanynone

Show simple item record