Technical Reference #107

107.

Fluorescence-Imaged Microdeformation Of The Outer Hair Cell Lateral Wall Oghalai; J.S.; Patel; A.A.; Nakagawa; T. and Brownell; W.E., Baylor College of Medicine, The Journal of Neuroscience, 18(107), (1998)
Link To Paper

Abstract:
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.

Microscopic Technique
Confocal Microscopy

Cell Type(s)
OTC