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Mechanotransducing (MS) ion channels and images of the patch membrane were studied in cell-attached patches in C6 glioma cells. MS channel density was approximately 0.08 to 0.5 channels/microns2, channel conductance was approximately 40 pS (at -40 mV), and the reversal potential was +15 mV. Replacement of NaCl with KCl, CsCl, or Na gluconate in the pipette solution was without substantial effect on the current-voltage relationship. Replacement of NaCl with NMDG (N-Methyl-D-Glucamine) Cl or reducing NaCl decreased the amplitude of inward currents at negative membrane potentials and caused the reversal potential to shift in the negative direction. Rapid application of suction to the back of the pipette usually elicited a fast (0.1 s) appearance of channel activity. The peak (phasic) in channel activity was followed by a decrease to a constant (tonic) level of activity. The reduction in channel activity--called adaptation--was reduced at depolarizing membrane potentials and disappeared if too much pressure was applied. Positive pressure caused the patch membrane to curve toward the pipette tip, move in the direction of the tip, and evoke MS channel activity. Removal of the positive pressure caused the patch to move back to the original position. Conversely, negative pressure caused the patch membrane to curve away from the pipette tip, move away from the tip, and elicit MS channel activity. Gigohm seal resistances were always maintained during translational movement of the patch membrane. Tonic MS channel activity was not associated with translational movements of the patch membrane. Phasic and tonic channel activity were independent of the sign of curvature of the patch membrane. C6 glioma cells have rapidly adapting voltage-dependent MS ion channels, which are non-selective for monovalent cations, and belong to the stretch-activating class of mechanosensory ion channels. Adaptation in MS channels may allow the cell to limit the influx of cations in response to mechanical input. The selective loss of adaptation suggests that the MS channel's gate receives input from two sources. A minimal viscoelastic mechanical model of adaptation and two alternative models for translational movement of the patch are presented. |
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