title: Ba2+ currents in inner and outer hair cells of mice lacking the voltage-dependent Ca2+ channel subunits beta3 or beta4.
abstract: Voltage-activated Ca(2+) channels comprise complexes of a pore-forming Ca(V)alpha(1) and auxiliary subunits Ca(V)beta, Ca(V)alpha(2)delta and sometimes Ca(V)gamma. The intracellular Ca(V)beta subunit assists in trafficking and surface expression of the Ca(V)alpha(1) subunit and can modulate biophysical properties of the Ca(2+) channel. Four genes, Ca(V)beta1-4, exist which confer different properties to Ca(2+) currents through the various Ca(V)alpha(1) subunits. Ca(2+) currents in cochlear inner (IHC) and outer hair cells (OHC) serving synaptic transmission flow predominantly through the L-type Ca(V)alpha(1) subunit Ca(V)1.3, but associated Ca(V)beta subunits are unknown. In the organ of Corti, we found mRNA and protein for all four Ca(V)beta subunits including Ca(V)beta2, but clear assignment of the Ca(V)beta1-4 immunolabelling with hair cells or nerve fibers was difficult. We analyzed Ca(V)beta3 knockout (Ca(V)beta3(-/-)) and Ca(V)beta4 mutant mice (Ca(V)beta4(lh/lh)), which had normal hearing. Recording voltage-activated Ba(2+) currents from hair cells of the two mouse models revealed distinct significant changes of cell size and Ba(2+) current properties compared with their wild-type controls. Neonatal Ca(V)beta4(lh/lh) IHCs showed reduced membrane capacitances and changes in the voltage dependence and kinetics of current activation, whereas mature IHCs had reduced peak currents compared with Ca(V)beta4(wt), altogether indicating the presence of Ca(V)beta4 in IHCs. Ba(2+) currents of Ca(V)beta3(-/-) OHCs showed largely reduced amplitudes, changes in the voltage dependence and kinetics of Ba(2+) current activation, and increased inactivation compared with Ca(V)beta3(wt), pointing to a role of Ca(V)beta3 for OHCs. These results indicate that neither Ca(V)beta3 nor Ca(V)beta4 are indispensable for hair cell Ca(2+) currents but contribute to the overall current properties.
authors.affiliation: Institute of Physiology II and Department of Otorhinolaryngology, Molecular Neurobiology, University of Tübingen, Tübingen Hearing Research Centre, Tübingen, Germany.
authors.forename: Stephanie
authors.initials: S
authors.lastname: Kuhn
authors.forename: Martina
authors.initials: M
authors.lastname: Knirsch
authors.forename: Lukas
authors.initials: L
authors.lastname: Rüttiger
authors.forename: Sylvia
authors.initials: S
authors.lastname: Kasperek
authors.forename: Harald
authors.initials: H
authors.lastname: Winter
authors.forename: Marc
authors.initials: M
authors.lastname: Freichel
authors.forename: Veit
authors.initials: V
authors.lastname: Flockerzi
authors.forename: Marlies
authors.initials: M
authors.lastname: Knipper
authors.forename: Jutta
authors.initials: J
authors.lastname: Engel
mesh_terms: D000818:Animals; D001464:Barium / Q000737:chemistry*; D002118:Calcium / Q000378:metabolism; D015220:Calcium Channels / Q000737:chemistry / Q000378:metabolism*; D003051:Cochlea / Q000378:metabolism; D004594:Electrophysiology / Q000379:methods; D004721:Endoplasmic Reticulum / Q000378:metabolism; D006198:Hair Cells, Auditory / Q000166:cytology / Q000378:metabolism*; D006309:Hearing; D007150:Immunohistochemistry / Q000379:methods; D007700:Kinetics; D051379:Mice; D051381:Rats; D017208:Rats, Wistar; D009435:Synaptic Transmission
chemical_list: C510225:Cacnb3 protein, mouse; C484333:Cacnb3 protein, rat; C491530:Cacnb4 protein, mouse; C491529:Cacnb4 protein, rat; D015220:Calcium Channels; D001464:Barium; D002118:Calcium