Patch-clamping drosophila sensory neurons

Volodymyr Kucher, Benjamin A Eaton, James D Stockand, Nina Boiko

Research output: Chapter in Book/Report/Conference proceedingChapter

1 Citation (Scopus)

Abstract

Electrophysiological studies provide essential clues about the regulation and physiological function of ion channel proteins. Probing ion channel activity in vivo, though, often is challenging. This can limit the usefulness of such model organisms as Drosophila for electrophysiological studies. This is unfortunate because these genetically tractable organisms represent powerful research tools that facilitate elaboration of complex questions of physiology. Here, we describe a recently developed method for recording ion channel activity in Drosophila sensory neurons. This approach is based on patch-clamping primary neuron cultures from Drosophila embryos. Such cultures allow the study of ion channels in different genetic backgrounds. In addition to describing how to prepare a primary neuronal cell culture from Drosophila embryos, we discuss, as an example of utility, analysis of Na+ currents in cultured class IV multidendritic (md) sensory neurons with the patch clamp technique. Excitability of md sensory neurons, manifested as action potential firing, is revealed with whole-cell current-clamping. Voltage-clamping class IV md neurons revealed the activity of the voltage-gated Na+ channel, paralytic. Moreover, challenging class IV md neurons with acidic pH activates acid-sensing inward Na+ currents. Genetic manipulation of Drosophila combined with this electrophysiological readout of activity identifies pickpocket1 (Ppk1), a member of the Deg/ENaC channel family, as responsible for conducting an acid-sensing Na+ current in class IV md sensory neurons.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
Pages385-397
Number of pages13
Volume998
DOIs
StatePublished - 2013

Publication series

NameMethods in Molecular Biology
Volume998
ISSN (Print)10643745

Fingerprint

Sensory Receptor Cells
Constriction
Drosophila
Ion Channels
Neurons
Embryonic Structures
Acids
Primary Cell Culture
Patch-Clamp Techniques
Action Potentials
Research
Proteins

Keywords

  • Drosophila sensory neurons
  • Ion channel
  • Patch clamping
  • Primary cell culture

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Cite this

Kucher, V., Eaton, B. A., Stockand, J. D., & Boiko, N. (2013). Patch-clamping drosophila sensory neurons. In Methods in Molecular Biology (Vol. 998, pp. 385-397). (Methods in Molecular Biology; Vol. 998). https://doi.org/10.1007/978-1-62703-351-0_30

Patch-clamping drosophila sensory neurons. / Kucher, Volodymyr; Eaton, Benjamin A; Stockand, James D; Boiko, Nina.

Methods in Molecular Biology. Vol. 998 2013. p. 385-397 (Methods in Molecular Biology; Vol. 998).

Research output: Chapter in Book/Report/Conference proceedingChapter

Kucher, V, Eaton, BA, Stockand, JD & Boiko, N 2013, Patch-clamping drosophila sensory neurons. in Methods in Molecular Biology. vol. 998, Methods in Molecular Biology, vol. 998, pp. 385-397. https://doi.org/10.1007/978-1-62703-351-0_30
Kucher V, Eaton BA, Stockand JD, Boiko N. Patch-clamping drosophila sensory neurons. In Methods in Molecular Biology. Vol. 998. 2013. p. 385-397. (Methods in Molecular Biology). https://doi.org/10.1007/978-1-62703-351-0_30
Kucher, Volodymyr ; Eaton, Benjamin A ; Stockand, James D ; Boiko, Nina. / Patch-clamping drosophila sensory neurons. Methods in Molecular Biology. Vol. 998 2013. pp. 385-397 (Methods in Molecular Biology).
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