Study of mechanisms of ionic transport and intracellular signaling pathways of shortening in cochlear outer hairce

点击翻译按钮获取中文摘要

The intracellular mechanisms of slow shortening in isolated guinea pig cochlear outer hair cells were investigated using inhibitors and/or an activator of protein kinases and protein phosphatases. The slow shortening was induced by tetanic electrical field stimulation.and changes in the cell length volume and intracellular CI^- concentration were microscopically monitored using a chloride-sensitive fluorescent dye. The slow shortening was inhibited by a calmodulin inhibitor. W-7, and a calcium calmodulin-dependent protein kinase II (CaMKII) inhibitor. KN-62. The inhibition by W-7 or KN-62, was abolished by the supplemented conductance of K^+ with valinomycin. Among the protein phosphatase inhibitors tested a type 1 and 2A protein phosphatase inhibitor. calyculin A.inhibited the slow shortening. The inhibition by calyculin A was abolished by the increased CI^- permeability. but neither by the increased K^+ conductance with valinomycin nor by the increased Ca^<2+> conductance with A23187. A protein serine/threonine phosphatase activator, N-acetylsphingosine. inhibited the shortening which was abolished by either valinomycin or a type 2A protein phosphatase inhibitor. okadaic acid but not by calyculin A.These findings suggest the following signaling mechanisms in the slow shortening of outer hair cells : the K^+ channel opening is facilitated through protein phosphorylation by CaMKII and suppressed via okadaic acid-sensitive dephosphorylation, and the C1^- channel opening depends on calyculin A-sensitive protein phosphatase activity.B.V.