On the effect of prestin on the electrical breakdown of cell membranes.
The voltage-dependent activity of prestin, the outer hair cell (OHC) motor protein essential for its electromotility, enhances the mammalian inner ear's auditory sensitivity. We investigated the effect of prestin's activity on the plasma membrane's (PM) susceptibility to electroporation (EP) via cell-attached patch-clamping. Guinea pig OHCs, TSA201 cells, and prestin-transfected TSA cells were subjected to incremental 50 mus and/or 50 ms voltage pulse trains, or ramps, at rates from 10 V/s to 1 kV/s, to a maximum transmembrane potential of +/-1000 mV. EP was determined by an increase in capacitance to whole-cell levels. OHCs were probed at the prestin-rich lateral PM or prestin-devoid basal portion; TSA cells were patched at random points. OHCs were consistently electroporated with 50 ms pulses, with significant resistance to depolarizing pulses. Although EP rarely occurred with 50 mus pulses, prior stimulation with this protocol had a significant effect on the sensitivity to EP with 50 ms pulses, regardless of polarity or PM domain. Consistent with these results, resistance to EP with depolarizing 10-V/s ramps was also found. Our findings with TSA cells were comparable, showing resistance to EP with both depolarizing 50-ms pulses and 10 V/s ramps. We conclude prestin significantly affects susceptibility to EP, possibly via known biophysical influences on specific membrane capacitance and/or membrane stiffness.
Navarrete
Enrique G
EG
Department of Cell and Molecular Biology, House Ear Institute, Los Angeles, California, USA.
Santos-Sacchi
Joseph
J
D000818:Animals; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism*; D004594:Electrophysiology; D018274:Electroporation; D006168:Guinea Pigs; D018072:Hair Cells, Auditory, Outer / Q000737:chemistry* / Q000378:metabolism; D006801:Humans; D008051:Lipid Bilayers / Q000737:chemistry; D009068:Movement; D018408:Patch-Clamp Techniques; D017434:Protein Structure, Tertiary; D011506:Proteins / Q000737:chemistry*; D013997:Time Factors; D014162:Transfection
D008051:Lipid Bilayers; C498722:Pres protein, Cavia porcellus; D011506:Proteins