Activation of mechanosensitive currents in traumatized membrane.

Mechanosensitive (MS) channels, ones whose open probability varies with membrane tension in patch recordings, are diverse and ubiquitous, yet many are remarkably insensitive to mechanical stimuli in situ. Failure to elicit mechanocurrents from cells with abundant MS channels suggests that, in situ, the channels are protected from mechanical stimuli. To establish what conditions affect MS channel gating, we monitored Lymnaea neuron stretch-activated K (SAK) channels in cell-attached patches after diverse treatments. Mechanosensitivity was gauged by rapidity of onset and extent of channel activation during a step pressure applied to a "naive" patch. The following treatments enhanced mechanosensitivity: actin depolymerization (cytochalasin B), N-ethylmaleimide, an inhibitor of ATPases including myosin, elevated Ca (using A-23187), and osmotic swelling (acutely and after 24 h). Osmotic shrinking decreased mechanosensitivity. A unifying interpretation is that traumatized cortical cytoskeleton cannot prevent transmission of mechanical stimuli to plasma membrane channels. Mechanoprotection and capricious mechanosensitivity are impediments to cloning efforts with MS channels. We demonstrate a potpourri of endogenous MS currents from L-M(TK-) fibroblasts; others had reported these cells to be MS current null and hence to be suitable for expressing putative MS channels.