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J Physiol Volume 557, Number 2, 379-388, June 1, 2004 DOI: 10.1113/jphysiol.2003.057687
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Patch clamp study of the UNC-105 degenerin and its interaction with the LET-2 collagen in Caenorhabditis elegans muscle

Maëlle Jospin1, Marie-Christine Mariol2, Laurent Segalat2 and Bruno Allard1

1 Physiologie Intégrative Cellulaire et Moléculaire, UMR CNRS 51232 Centre de Génétique Moléculaire et Cellulaire, UMR CNRS 5534, Université C. Bernard Lyon I, 43 bd du 11 Novembre 1918, 69622 Villeurbanne cedex, France

Degenerins have emerged from genetic studies in Caenorhabditis elegans as candidate mechanically gated amiloride-sensitive ion channels for transducing mechanical stimuli into cellular responses. In C. elegans muscle, the existence of a genetic interaction between the unc-105 degenerin gene and let-2, a gene encoding an {alpha}2(IV) collagen, raised the possibility that UNC-105 may function as a mechanically gated channel in a stretch receptor complex. However, to date, ion channel activity of UNC-105 has only been recorded in a gain-of-function mutant form in heterologous expression systems. In this study we investigated the in situ properties of UNC-105 using the whole cell configuration of the patch clamp technique on body wall muscle cells from acutely dissected C. elegans. Amiloride was found to be without effect on membrane potential of wild-type muscle cells, suggesting that the UNC-105 degenerin is electrically silent in resting muscle. Hypo-osmotic shocks induced a reversible depolarization of muscle cells but which was not affected by amiloride. Deformation of the cells by applying tension to the filamentous complex on which muscle cells remained attached or by ejecting external solution under pressure failed to induce any change of membrane potential. In gain-of-function unc-105(n506) mutant cells, an amiloride-sensitive inward Na+ current was found to be constitutively active, leading to maintained muscle depolarization. An associated mutation in the {alpha}2(IV) collagen LET-2 led to the closure of the mutant UNC-105(n506) channel while a collagenase treatment of these double mutant cells caused it to re-open, giving evidence for a functional interaction between LET-2 collagen and mutant UNC-105 channel.

(Received 5 November 2003; accepted after revision 8 March 2004; first published online 12 March 2004)
Corresponding author B. Allard: Physiologie Intégrative Cellulaire et Moléculaire, UMR CNRS 5123, University Claude Bernard Lyon 1, 43 bd 11 Novembre 1918, 69622 Villeurbanne cedex, France. Email: bruno.allard{at}univ-lyon1.fr




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