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The Journal of Neuroscience, January 1, 1998, 18(1):48-58

Fluorescence-Imaged Microdeformation of the Outer Hair Cell Lateral Wall

John S. Oghalai¹, Alpen A. Patel¹, Takashi Nakagawa^{1, 2}, and William E. Brownell¹

¹ Bobby R. Alford Department of Otorhinolaryngology and Communicative Sciences, Baylor College of Medicine, Houston, Texas 77030, and ² Department of Otorhinolaryngology, Faculty of Medicine, Kyushu University, Fukuoka 812, Japan

Outer hair cell (OHC) electromotility appears to be central to mammalian hearing and originates within its lateral wall. The OHC lateral wall is a unique trilaminate structure consisting of the plasma membrane (PM), the cortical lattice (CL), and the subsurface cisternae (SSC). We selectively labeled and imaged the lateral wall components in the isolated guinea pig OHC under confocal microscopy. The PM was labeled with a voltage-sensitive dye, di-8-ANEPPS; the SSC was labeled with the sphingomyelin precursor, NBD-C₆-ceramide; and F-actin in the CL was labeled with conjugates of phalloidin. Interactions among the three layers were evaluated with the micropipette aspiration technique. The PM was tethered to the CL and SSC until, at a critical deformation pressure, the PM separated, allowing visualization of the extracisternal space, and ultimately formed a vesicle. After detaching, the stiffness parameter of the PM was 22% of that of the intact lateral wall. We conclude that the lateral wall PM is more compliant than the CL/SSC complex. The data clarify the structural basis for electromotile force coupling in the OHC lateral wall.

Key words: cochlea; inner ear; hearing; cytoskeleton; cell stiffness; stiffness parameter; micropipette aspiration; patch-clamp technique; confocal microscopy; fluorescence labeling; biophysics

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Copyright © 1998 Society for Neuroscience  0270-6474/98/18148-11$05.00/0

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