The Journal of Neuroscience, June 18, 2008, 28(25):6342-6353; doi:10.1523/JNEUROSCI.1154-08.2008
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Cellular/Molecular
The Dimensions and Composition of Stereociliary Rootlets in Mammalian Cochlear Hair Cells: Comparison between High- and Low-Frequency Cells and Evidence for a Connection to the Lateral Membrane
David N. Furness,1
Shanthini Mahendrasingam,1
Mitsuru Ohashi,1
Robert Fettiplace,2 and
Carole M. Hackney3
1Institute of Science and Technology in Medicine, Keele University, Keele, Staffordshire ST5 5BG, United Kingdom, 2Department of Physiology, University of Wisconsin Medical School, Madison, Wisconsin 53706, and 3Department of Physiology, Development, and Neuroscience, University of Cambridge, Cambridge CB2 3EG, United Kingdom
Correspondence should be addressed to David N. Furness, Institute of Science and Technology in Medicine, Keele University, Keele, Staffordshire ST5 5BG, UK. Email: coa14{at}keele.ac.uk
The sensory bundle of vertebrate cochlear hair cells consists of actin-containing stereocilia that are thought to bend at their ankle during mechanical stimulation. Stereocilia have dense rootlets that extend through the ankle region to anchor them into the cuticular plate. Because this region may be important in bundle stiffness and durability during prolonged stimulation at high frequencies, we investigated the structure and dimensions of rootlets relative to the stereocilia in apical (low-frequency) and basal (high-frequency) regions of rodent cochleae using light and electron microscopy. Their composition was investigated using postembedding immunogold labeling of tropomyosin, spectrin, β-actin,
-actin, espin, and prestin. The rootlets have a thick central core that widens at the ankle, and are embedded in a filamentous meshwork in the cuticular plate. Within a particular frequency region, rootlet length correlates with stereociliary height but between regions it changes disproportionately; apical stereocilia are, thus, approximately twice the height of basal stereocilia in equivalent rows, but rootlet lengths increase much less. Some rootlets contact the tight junctions that underlie the ends of the bundle. Rootlets contain spectrin, tropomyosin, and β- and
-actin, but espin was not detected; spectrin is also evident near the apical and junctional membranes, whereas prestin is confined to the basolateral membrane below the junctions. These data suggest that rootlets strengthen the ankle region to provide durability and may contact with the lateral wall either to give additional anchoring of the stereocilia or to provide a route for interactions between the bundle and the lateral wall.
Key words: cochlea; hair cells; stereocilia; stereociliary rootlets; stereociliary stiffness; cuticular plate
Received March 17, 2008;
revised May 7, 2008;
accepted May 7, 2008.
Correspondence should be addressed to David N. Furness, Institute of Science and Technology in Medicine, Keele University, Keele, Staffordshire ST5 5BG, UK. Email: coa14{at}keele.ac.uk
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