Regulation of Ca2+ -dependent K+ channel expression in rat cerebellum during postnatal development

Abstract

Potassium channels govern duration and frequency of excitable membrane events and may regulate signals that are important in neuronal development. This study assesses the developmental expression of the large conductance Ca2+ -dependent K+ channel in vivo and in vitro in rat cerebellum. In vivo, transcript levels for the Ca2+ -dependent K+ channel (K(Ca)) were shown by Northern analysis to increase during development, whereas transcript levels for the voltage-gated K+ channel Kv3.1, a delayed rectifier (K(D)), remained relatively constant. A comparable pattern was demonstrated by expression in Xenopus oocytes of poly(A)-enriched RNA isolated from postnatal rat cerebella. In cerebellar cultures, increased external K+ provided a simple manipulation of cell excitability that influenced K(Ca) transcript levels during development. With low external K+ (5.3 mM), the levels of K(Ca) channel transcript (assessed by semiquantitative PCR) remained constant throughout development. However, in culture medium that supported significant dendritic outgrowth (10 mM extracellular K+), an upregulation of K(Ca) transcript level was observed similar to that seen in vivo. Tetraethylammonium (TEA; 1 mM) similarly enhanced K(Ca) expression, suggesting that depolarizing stimuli increased K(Ca) expression. The stimulatory effects of increased K+ or TEA on K(Ca) expression required extracellular Ca2+ and were abolished in low external calcium (0.1 mM, buffered with EGTA), although morphological development and survival were not impaired. The regulation of K(Ca) channel expression by depolarization and Ca2+ entry provides evidence of a logical feedback mechanism governing Ca2+ signals that may be significant in cerebellar development.