Cytologic evidence for mechanisms of K+ transport and genesis of Hensen bodies and subsurface cisternae in outer hair cells.
A system commonly termed the tubulocisternal endoplasmic reticulum (TCER), but designated here the canalicular reticulum (CR), occurs selectively in ion-transporting epithelia, in which it is interpreted as facilitating the transcellular diffusion of ions. Mechanoelectrical transduction in the cochlear outer hair cells (OHCs) depends on the apical influx and the subsequent basolateral efflux of K+. Cytologic structures that possibly mediate K+ transport in gerbil OHCs were investigated here.



Cochleas were fixed primarily with glutaraldehyde and secondarily with reagents for demonstrating TCER or were fixed with a ferrocyanide-osmium tetroxide solution to preserve intracellular membranes. The distribution of membranous structures retained with these techniques was examined by using electron microscopy.



Secondary fixation with osmium tetroxide-ferrocyanide permitted ultrastructural demonstration in OHCs of increased numbers of Hensen bodies and newly detected membranous systems, including CR, linear cisternae, small clusters of cytosolic vesicles and complexes of canaliculi, segmented cisternae, and mitochondria. CR filling an apical stratum beside and below the cuticular plate and contacting laterally the uppermost subsurface cisternae (SSC) was situated to sequester and transport the apical K+ influx that attends the acoustically generated receptor potential and the silent current. The close association of CR with numerous, highly developed Golgi bodies exclusively in the apex of the cell suggested genesis of CR from Golgi cisternae. Nonbranching, linear cisternae occupied a lower cell stratum and spread from CR laterally to a more inferior region of the SSC. Small clusters of vesicles in the central cytosol resembled Hensen bodies in their envelopment by branching canaliculi and segmented cisternae in close association with mitochondria. Viewing the vesicles in Hensen bodies and the small clusters as functioning like most other cytoplasmic vesicles in transport of cell membrane permitted the interpretation that these vesicles move nascent membrane from the canalicular-mitochondrial complex to the SSC. Other small clusters of vesicles contacted the innermost layer of the SSC, often at cisterna-depleted foci in which the vesicles appeared to either replenish the SSC or arise in the course of its turnover. Proximity of multivesicular bodies and lysosomes to small vesicle clusters in foci of depleted SSC implicated the lysosomes in digesting vesicles released from the SCC. Populations of unique, large, lysosome-like bodies and of small, dense bodies in the upper cytosol of OHCs appeared to be involved in different catabolic pathways mediating the turnover ofSSC, CR, and other structures.



Cochlear OHCs contain previously unrecognized membranous organelles that facilitate ion transport and presumably contribute thereby to mechanoelectrical transduction. Vesicles in small clusters and Hensen bodies arise from complexes of canaliculi, cisternae, and mitochondria and contribute membrane to the genesis of the SSC.
Spicer
S S
SS
Department of Pathology and Laboratory Medicine, Medical University of South Carolina, Charleston 29425, USA.
Thomopoulos
G N
GN
Schulte
B A
BA
D000818:Animals; D003051:Cochlea / Q000166:cytology / Q000378:metabolism / Q000648:ultrastructure; D003594:Cytoplasmic Granules / Q000378:metabolism / Q000648:ultrastructure; D003600:Cytosol / Q000737:chemistry / Q000378:metabolism / Q000648:ultrastructure; D004721:Endoplasmic Reticulum / Q000378:metabolism / Q000648:ultrastructure; D005849:Gerbillinae; D018072:Hair Cells, Auditory, Outer / Q000166:cytology / Q000378:metabolism / Q000648:ultrastructure*; D007425:Intracellular Membranes / Q000378:metabolism / Q000648:ultrastructure; D017136:Ion Transport; D008854:Microscopy, Electron; D008928:Mitochondria / Q000378:metabolism / Q000648:ultrastructure; D011188:Potassium / Q000378:metabolism*
D011188:Potassium