Show simple item record

dc.contributor.authorKatz, Ben
dc.contributor.authorGutorov, Ren
dc.contributor.authorHardie, Rogeren
dc.contributor.authorMinke, Ben
dc.date.accessioned2017-06-22T11:31:41Z
dc.date.available2017-06-22T11:31:41Z
dc.date.issued2017-06-13en
dc.identifier.issn1940-087X
dc.identifier.urihttps://www.repository.cam.ac.uk/handle/1810/264968
dc.description.abstractWhole-cell voltage clamp recordings from $\textit{Drosophila melanogaster}$ photoreceptors have revolutionized the field of invertebrate visual transduction, enabling the use of $\textit{D. melanogaster}$ molecular genetics to study inositol-lipid signaling and Transient Receptor Potential (TRP) channels at the single-molecule level. A handful of labs have mastered this powerful technique, which enables the analysis of the physiological responses to light under highly controlled conditions. This technique allows control over the intracellular and extracellular media; the membrane voltage; and the fast application of pharmacological compounds, such as a variety of ionic or pH indicators, to the intra- and extracellular media. With an exceptionally high signal-to-noise ratio, this method enables the measurement of dark spontaneous and light-induced unitary currents ( i.e. spontaneous and quantum bumps) and macroscopic Light-induced Currents (LIC) from single $\textit{D. melanogaster}$ photoreceptors. This protocol outlines, in great detail, all the key steps necessary to perform this technique, which includes both electrophysiological and optical recordings. The fly retina dissection procedure for the attainment of intact and viable $\textit{ex vivo}$ isolated ommatidia in the bath chamber is described. The equipment needed to perform whole-cell and fluorescence imaging measurements are also detailed. Finally, the pitfalls in using this delicate preparation during extended experiments are explained.
dc.description.sponsorshipThe experimental part of this research was supported by grants from the US-Israel Bi National Science Foundation (to B.M. and I.L.), the Israel Science Foundation (ISF), the Deutsch-Israelische Projektkooperation (DIP) (to B.M.), and the Biotechnology and Biological Sciences Research Council (BBSRC Grant numbers: BB/M007006/1 and BB/D007585/1) to R.C.H.
dc.languageengen
dc.language.isoenen
dc.publisherJoVE
dc.rightsAttribution 4.0 Internationalen
dc.rightsAttribution 4.0 Internationalen
dc.rightsAttribution 4.0 Internationalen
dc.rightsAttribution 4.0 Internationalen
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/en
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/en
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/en
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/en
dc.subjectneuroscienceen
dc.subjectissue 12en
dc.subjectwhole-cell recordingen
dc.subjectvoltage clamen
dc.subjectcurrent clamen
dc.subjectelectrophysiologyen
dc.subjectDrosophila melanogasteren
dc.subjectphototransductionen
dc.subjectsingle-cell Ca 2+ imagingen
dc.subjectintracellular perfusionen
dc.titleElectrophysiological Method for Whole-cell Voltage Clamp Recordings from $\textit{Drosophila}$ Photoreceptorsen
dc.typeArticle
prism.numbere55627en
prism.publicationDate2017en
prism.publicationNameJournal of Visualized Experimentsen
prism.volume124en
dc.identifier.doi10.17863/CAM.10824
dcterms.dateAccepted2017-01-24en
rioxxterms.versionofrecord10.3791/55627en
rioxxterms.versionVoRen
rioxxterms.licenseref.urihttp://creativecommons.org/licenses/by/4.0/en
rioxxterms.licenseref.startdate2017-06-13en
dc.contributor.orcidHardie, Roger [0000-0001-5531-3264]
dc.identifier.eissn1940-087X
rioxxterms.typeJournal Article/Reviewen
pubs.funder-project-idBBSRC (BB/D007585/1)
pubs.funder-project-idBBSRC (BB/M007006/1)
cam.orpheus.successThu Jan 30 12:53:51 GMT 2020 - The item has an open VoR version.*
rioxxterms.freetoread.startdate2100-01-01


Files in this item

Thumbnail
Thumbnail

This item appears in the following Collection(s)

Show simple item record

Attribution 4.0 International
Except where otherwise noted, this item's licence is described as Attribution 4.0 International