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Related Experiment Videos

Single-channel currents through transient-receptor-potential-like (TRPL) channels.

J Hambrecht1, S Zimmer, V Flockerzi

  • 1Institut für Pharmakologie und Toxikologie, Universität des Saarlandes, Homburg/Saar, Germany.

Pflugers Archiv : European Journal of Physiology
|August 23, 2000
PubMed
Summary

Drosophila transient-receptor-potential-like (TRPL) channels are constitutively active, non-selective cation channels. Intracellular calcium and membrane depolarization modulate TRPL channel activity in mammalian cells.

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Area of Science:

  • Molecular Biology
  • Ion Channel Physiology
  • Cellular Electrophysiology

Background:

  • Transient Receptor Potential (TRP) channels are crucial for cellular signaling.
  • Drosophila TRPL channels, a subtype of TRP channels, have known roles in insects.
  • Understanding TRPL channel properties in mammalian cells is essential for comparative physiology.

Purpose of the Study:

  • To characterize the biophysical properties of Drosophila TRPL channels expressed in mammalian cells.
  • To investigate the modulation of TRPL channel activity by membrane potential and intracellular ions.
  • To determine if TRPL channels can function and be modulated in a heterologous mammalian system.

Main Methods:

  • Single-channel current recordings in HEK and COS cells expressing recombinant TRPL channels.

Related Experiment Videos

  • On-cell and inside-out patch-clamp electrophysiology.
  • Varying ionic concentrations (Na+, Mg2+, Ca2+, Ba2+) and intracellular guanosine 5'-O-(3-thiotriphosphate) (GTP[gamma-S]) and Ca2+ levels.

    • Main Results:

    • TRPL channels exhibited constitutive activity with specific single-channel conductances (e.g., 104 pS with 115 mM Na+).
    • Channel currents reversed near 0 mV, showing linear voltage dependence and cation selectivity.
    • Opening probability increased with depolarization, causing outward rectification; activity was enhanced by GTP[gamma-S] and elevated intracellular Ca2+ (>2 microM).
    • Constitutively active TRPL channels depolarized host cells, which was reversed by BAPTA chelation.

    Conclusions:

    • Drosophila TRPL channels expressed in mammalian cells are constitutively active, non-selective cation channels.
    • TRPL channel function is modulated by membrane potential and intracellular calcium levels.
    • These findings suggest a conserved role and modulation mechanism for TRPL channels across species.