Imaging of the cortical cytoskeleton of guinea pig outer hair cells using atomic force microscopy

Mammalian outer hair cells (OHCs) are known to respond to acoustical stimulation with elongation and contraction of the cells' cylindrical soma in vivo, and this motility is related to both the protein motors distributed along the OHC plasma membrane and the cytoskeleton beneath it. Therefore, the cytoskeleton seems to play an important role in the motility of the OHC. Recently, an atomic force microscope (AFM) was used to investigate the OHC cytoskeleton under physiological conditions. However, details were not made clear in that study. In this study, the ultrastructure of the cytoskeleton of fixed OHCs of guinea pigs, which were extracted with Triton X-100, was investigated using the AFM. As a result, the cortical cytoskeleton, which is formed by discrete oriented domains, was imaged, and circumferential filaments and cross-links were observed within the domain. Morphological change of the cytoskeleton of the OHC induced by diamide treatment was then examined using the AFM, and reduction of cross-links was observed. The examination indicates that the cortical cytoskeleton comprises circumferential actin filaments and spectrin cross-links.