Electrophysiological method for whole-cell voltage clamp recordings from drosophila photoreceptors

Ben Katz, Rita Gutorov, Elisheva Rhodes-Mordov, Roger C. Hardie*, Baruch Minke

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

6 Scopus citations

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.

Original languageEnglish
Article numbere55627
JournalJournal of Visualized Experiments
Volume2017
Issue number124
DOIs
StatePublished - 13 Jun 2017

Bibliographical note

Publisher Copyright:
© 2017 Journal of Visualized Experiments.

Keywords

  • Current clamp
  • Drosophila melanogaster
  • Electrophysiology
  • Intracellular perfusion
  • Issue 124
  • Neuroscience
  • Phototransduction
  • Single-cell Ca imaging
  • Voltage clamp
  • Whole-cell recordings

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