Transport Processes, Metabolism and Endocrinology; Kidney, Gastrointestinal Tract, and Exocrine Glands

Pflügers Archiv

, Volume 421, Issue 6, pp 606-612

First online:

Pressure-clamp: a method for rapid step perturbation of mechanosensitive channels

  • Don W. McBrideJr.Affiliated withSection of Neurobiology and Behavior, Cornell University
  • , Owen P. HamillAffiliated withSection of Neurobiology and Behavior, Cornell University

Rent the article at a discount

Rent now

* Final gross prices may vary according to local VAT.

Get Access

Abstract

Here we describe a pressure-clamp method for applying suction or pressure steps to membrane patches in order to study the activation, adaptation and relaxation characteristics of mechanosensitive (MS) channels. A description is given of the mechanical arrangement of the pressure clamp which involves a balance between negative (suction) and positive pressures. The electronic circuitry of the feedback control is described. We also describe the optimal time response (≈ 10 ms) of the pressure-clamp, the amplitude of pressure resolution (0.2–0.5 mmHg; 27–67 Pa) and the factors influencing these parameters. We illustrate the applications of the clamp on the Xenopus oocyte and cultured skeletal myotubes from dystrophic mouse (mdx) muscle, both of which express MS channels. Studies with pressure/suction pulses indicate that in both muscle and oocytes MS channel activity displays adaptation. The ability to study current relaxations following step changes in pressure/suction using the pressure-clamp in combination with patch-clamp techniques provides the opportunity for analysis of the time, voltage and pressure dependence of the opening and closing of MS channels.

Key words

Pressure-clamp Mechanosensitive channels Rapid mechanical perturbation Xenopus oocytes Dystrophic muscle