Amphipath-induced nanoscale changes in outer hair cell plasma membrane curvature.
Outer hair cell (OHC) electromotility enables frequency selectivity and sensitivity in mammalian audition. Electromotility is generated by the transmembrane protein prestin and is sensitive to amphipathic compounds including salicylate, chlorpromazine (CPZ), and trinitrophenol (TNP). Although these compounds induce observable membrane curvature changes in erythrocytes, their effects on OHC membrane curvature are unknown. In this work, fluorescence polarization microscopy was applied to investigate the effects of salicylate, CPZ, and TNP on di-8-ANEPPS orientation in the OHC plasma membrane. Our results demonstrate the ability of fluorescence polarization microscopy to measure amphipath-induced changes in di-8-ANEPPS orientation, consistent with nanoscale changes in membrane curvature between regularly spaced proteins connecting the OHC plasma membrane and cytoskeleton. Simultaneous application of oppositely charged amphipaths generally results in no net membrane bending, consistent with predictions of the bilayer couple hypothesis; however, the application of salicylate (10 mM), which inhibits electromotility, is not reversed by the addition of CPZ. This result supports other findings that suggest salicylate primarily influences electromotiliy and OHC nonlinear capacitance via a direct interaction with prestin. In contrast, we find that CPZ and TNP influence the voltage sensitivity of prestin via membrane bending, demonstrating the mechanosensitivity of this unique membrane motor protein.
Greeson
Jennifer N
JN
Rice University, Department of Bioengineering, Houston, Texas 77251-1892, USA.
Raphael
Robert M
RM
D000704:Analysis of Variance; D000818:Animals; D002462:Cell Membrane / Q000187:drug effects* / Q000378:metabolism / Q000648:ultrastructure*; D002746:Chlorpromazine / Q000494:pharmacology; D003599:Cytoskeleton / Q000187:drug effects / Q000378:metabolism; D005260:Female; D006168:Guinea Pigs; D018072:Hair Cells, Auditory, Outer / Q000187:drug effects* / Q000378:metabolism / Q000648:ultrastructure*; D008564:Membrane Potentials; D008856:Microscopy, Fluorescence; D010851:Picrates / Q000494:pharmacology; D011506:Proteins / Q000187:drug effects / Q000378:metabolism; D011726:Pyridinium Compounds; D012459:Salicylates / Q000494:pharmacology; D013501:Surface-Active Agents / Q000494:pharmacology*
C089669:1-(3-sulfonatopropyl)-4-(beta-(2-(di-n-octylamino)-6-naphthyl)vinyl)pyridinium betaine; D010851:Picrates; D011506:Proteins; D011726:Pyridinium Compounds; D012459:Salicylates; D013501:Surface-Active Agents; C005858:picric acid; D002746:Chlorpromazine