A genetically targetable fluorescent probe of channel gating with rapid kinetics

Kazuto Ataka, Vincent A. Pieribone

Research output: Contribution to journalArticle

147 Citations (Scopus)

Abstract

We have developed a genetically targetable, optical channel-gating reporter that converts rapid membrane potential changes into changes in fluorescence intensity. We have named this construct SPARC (sodium channel protein-based activity reporting construct). Green fluorescent protein was inserted into an intracellular loop of a reversibly nonconducting form of the rat μl skeletal muscle voltage-gated sodium channel. Rapid changes of the membrane potential modulate the fluorescence of the inserted green fluorescent protein. This change in fluorescence can faithfully report depolarizing pulses as short as 2 ms. The fluorescence signal does not inactivate during extended depolarizations. Several features of the probe's response properties indicate that it likely reports gating charge movement of a single domain of rat μl skeletal muscle. This probe provides a new approach for studying rapid channel movements and may possibly act as a fluorescent activity reporter in excitable cells.

Original languageEnglish
Pages (from-to)509-516
Number of pages8
JournalBiophysical Journal
Volume82
Issue number1
DOIs
Publication statusPublished - 2002

ASJC Scopus subject areas

  • Biophysics

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