Antisense knockdown of calcium-dependent K + channels in developing cerebellar Purkinje neurons

• Published in: Brain research. Developmental brain research 2000-04, Vol.120 (2), p.135-140
• Main Authors: Muller, Yunhua Li, Reitstetter, Raven, Yool, Andrea J
• Format: Article
• Language: English
• Subjects: Animals; Antisense oligonucleotide; Apamin - pharmacology; BK channel; Calbindins; Cell Differentiation - drug effects; Cell Differentiation - genetics; Cerebellar Cortex - cytology; Cerebellar Cortex - drug effects; Cerebellar Cortex - embryology; Down-Regulation - drug effects; Down-Regulation - genetics; Fetus; Gene Expression Regulation, Developmental - drug effects; Gene Expression Regulation, Developmental - physiology; Immunohistochemistry; Membrane Potentials - drug effects; Membrane Potentials - genetics; Oligonucleotides, Antisense - pharmacology; Patch clamp electrophysiology; Potassium Channels, Calcium-Activated - drug effects; Potassium Channels, Calcium-Activated - genetics; Potassium Channels, Calcium-Activated - metabolism; Purkinje Cells - cytology; Purkinje Cells - drug effects; Purkinje Cells - metabolism; Rats; rslo; S100 Calcium Binding Protein G - metabolism; Transcription, Genetic - drug effects; Transcription, Genetic - physiology
• ISSN: 0165-3806
• DOI: 10.1016/S0165-3806(00)00004-3

Normal developmental upregulation of K Ca channel activity in cultured rat cerebellar Purkinje neurons was selectively inhibited by antisense oligonucleotide sequence (3 μM) targeted against the rslo transcript. The knockdown was specific; delayed rectifier and apamin-sensitive K + channel abundances in Purkinje neurons were not affected by rslo antisense. Sense oligonucleotides (3 μM), used as a control, had no effect on channel abundance. Quantitative morphometric analyses of anti-calbindin-labeled Purkinje neurons showed no differences between neurons in control, sense and antisense treatment groups, and confirmed that the presence of the added oligonucleotide in the sense and antisense treatment conditions had no discernable toxic effects on neuronal health, for which neurite outgrowth is a sensitive indicator. These results confirm the identification of the developmentally regulated K Ca channel as the product of the gene rslo in cerebellar Purkinje neurons.