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A-Type K+ Current Mediated by the Kv4 Channel Regulates the Generation of Action Potential in Developing Cerebellar Granule Cells
Riichi Shibata1, 3, Kensuke Nakahira1, 4, Koji Shibasaki1, 3, Yoshihiko Wakazono1, Keiji Imoto2, 3, and Kazuhiro Ikenaka1, 3 1 Laboratory of Neural Information, 2 Laboratory of Humoral Information, Department of Informational Physiology, and 3 Department of Physiological Sciences, the Graduate University for Advanced Studies, National Institute for Physiological Sciences, and 4 Center for Bio-Environmental Research, National Institute for Basic Biology, Okazaki National Research Institutes, Okazaki, Aichi 444-8585, Japan
During neuronal differentiation and maturation, electrical excitability is essential for proper gene expression and the formation of synapses. The expression of ion channels is crucial for this process; in particular, voltage-gated K+ channels function as the key determinants of membrane excitability. Previously, we reported that the A-type K+ current (IA) and Kv4.2 K+ channel subunit expression increased in cultured cerebellar granule cells with time. To examine the correlation between ion currents and the action potential, in the present study, we measured developmental changes of action potentials in cultured granule cells using the whole-cell patch-clamp method. In addition to an observed increment of IA, we found that the Na+ current also increased during development. The increase in both currents was accompanied by a change in the membrane excitability from the nonspiking type to the repetitive firing type. Next, to elucidate whether Kv4.2 is responsible for the IA and to assess the effect of Kv4 subunits on action potential waveform, we transfected a cDNA encoding a dominant-negative mutant Kv4.2 (Kv4.2dn) into cultured cells. Expression of Kv4.2dn resulted in the elimination of IA in the granule cells. This result demonstrates that members of the Kv4 subfamily are responsible for the IA in developing granule cells. Moreover, elimination of IA resulted in shortening of latency before the first spike generation. In contrast, expression of wild-type Kv4.2 resulted in a delay in latency. This indicates that appearance of IA is critically required for suppression of the excitability of granule cells during their maturation.
Key words: Kv4.2; A-type current; dominant-negative; transfection; action potential; fast spike latency; microexplant culture; whole-cell patch clamp
Copyright © 2000 Society for Neuroscience 0270-6474/00/20114145-11$05.00/0
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