Sub-millisecond ligand probing of cell receptors with multiple solution exchange

Sergiy Sylantyev*, Dmitri A. Rusakov

*Corresponding author for this work

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

The accurate knowledge of receptor kinetics is crucial to our understanding of cell signal transduction in general and neural function in particular. The classical technique of probing membrane receptors on a millisecond scale involves placing a recording micropipette with a membrane patch in front of a double-barrel (θ-glass) application pipette mounted on a piezo actuator. Driven by electric pulses, the actuator can rapidly shift the θ-glass pipette tip, thus exposing the target receptors to alternating ligand solutions. However, membrane patches survive for only a few minutes, thus normally restricting such experiments to a single-application protocol. In order to overcome this deficiency, we have introduced pressurized supply microcircuits in the θ-glass channels, thus enabling repeated replacement of application solutions within 10–15 s. This protocol, which has been validated in our recent studies and takes 20–60 min to implement, allows the characterization of ligand-receptor interactions with high sensitivity, thereby also enabling a powerful paired-sample statistical design.
Original languageEnglish
Pages (from-to)1299–1306
Number of pages8
JournalNature Protocols
Volume8
Issue number7
Early online date6 Jun 2013
DOIs
Publication statusPublished - Jul 2013

Keywords

  • Molecular neuroscience
  • Patch clamp
  • Receptor pharmacology

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