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KMID : 0361020090520080655
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Korean Journal of Otolaryngology - Head and Neck Surgery
2009 Volume.52 No. 8 p.655 ~ p.659
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Isolation of Voltage Dependent Calcium Current in Chick Inner Hair Cell
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Lee Seung-Hwan
Shin Myung-Chul
Ahn Tae-Hwan
Kim Ki-Yong
Kim Kuk
Min Hyun-Jung
Park Chul-Won
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Abstract
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Background and Objectives : Voltage dependent calcium channel (VDCC) mediates calcium ion influx and controls neurotransmitter release in excitable cells. Hair cells in vertebrates cochlea are known to express L-type VDCC. The purpose of this study was to measure calcium current from hair cells to investigate basic activity and characteristics of VDCC.
Materials and Method : We measured calcium current in hair cells of the chicken’s auditory organ, the basilar papilla analogous to the mammalian cochlea, in whose L-type, dihydropyridinesensitive calcium channels predominate and in vestibular hair cells from cristae. Calcium current through VDCC was isolated in voltage-clamp recording using Cesium, Tetraethylammonium, 4- aminopyridine and apamin to block the much larger potassium currents. Various concentrations of internal calcium buffer, ethylene glycol tetraacetic acid (EGTA) or 1,2-bis (o-aminophenoxy) ethane-N,N,N’,N’-tetraacetic acid (BAPTA) were used.
Results:The higher the buffer concentration, the larger the current size were; they were significantly larger in 10 mM of calcium buffer concentration (ANOVA, p<0.05). There was no difference in calcium current between cochlear and vestibular hair cells.
Conclusion: We could successfully isolate stable inward calcium current from chick hair cells. This experiment can be used as a basic method to understand neurotransmission process between hair cells and afferent neurons.
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KEYWORD
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Hair cell, Calcium channel
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