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Electrophysiological Method for Whole-cell Voltage Clamp Recordings from $\textit{Drosophila}$ Photoreceptors

Published version
Peer-reviewed

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Authors

Katz, B 
Gutorov, R 
Hardie, RC 
Minke, B 

Abstract

Whole-cell voltage clamp recordings from Drosophila melanogaster photoreceptors have revolutionized the field of invertebrate visual transduction, enabling the use of 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 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 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.

Description

Keywords

neuroscience, issue 12, whole-cell recording, voltage clam, current clam, electrophysiology, Drosophila melanogaster, phototransduction, single-cell Ca 2+ imaging, intracellular perfusion

Journal Title

Journal of Visualized Experiments

Conference Name

Journal ISSN

1940-087X
1940-087X

Volume Title

124

Publisher

JoVE
Sponsorship
Biotechnology and Biological Sciences Research Council (BB/D007585/1)
Biotechnology and Biological Sciences Research Council (BB/M007006/1)
The 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.