Organization of membrane motor in outer hair cells: an atomic force microscopic study

Using atomic force microscopy, we imaged the cytosolic surface of the lateral plasma membrane of outer hair cells from guinea pigs' inner ear. We used a "cell-free" preparation, in which a patch of plasma membrane was firmly attached to a substrate and the cytoplasmic face was exposed. The membrane patches contained densely packed particles whose diameter, after correcting for the geometry of the probing tip, was approximately 10 nm. The particles were predominantly aligned unidirectionally with spacing of approximately 36 nm. The density of the particle was approximately 850 microm(-2), which could be an underestimate presumably due to the method of sample preparation. Antibody-labeled specimens showed particles more elevated than unlabeled preparation indicative of primary and secondary antibody complexes. The corrected diameters of these particles labeled with anti-actin were approximately 12 nm while that with antiprestin were approximately 8 nm. The alignment pattern in antiprestin-labeled specimens resembled that of the unlabeled preparation. Specimens labeled with actin antibodies did not show such alignment. We interpret that the particles observed in the unlabeled membranes correspond to the 10-nm particles reported by electron microscopy and that these particles contain prestin, a member of the SLC26 family, which is essential for electromotility.