A role for inwardly rectifying K+ channels in differentiation of NG108-15 neuroblastoma x glioma cells
| Autor: | J J, Pancrazio, W, Ma, G M, Grant, K M, Shaffer, W Y, Kao, Q Y, Liu, P, Manos, J L, Barker, D A, Stenger |
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| Rok vydání: | 1999 |
| Předmět: |
Neurons
ERG1 Potassium Channel Patch-Clamp Techniques Potassium Channels Reverse Transcriptase Polymerase Chain Reaction Cesium Cell Differentiation Glioma Hybrid Cells Ether-A-Go-Go Potassium Channels Choline O-Acetyltransferase Membrane Potentials Rats DNA-Binding Proteins Mice Neuroblastoma Chlorides Transcriptional Regulator ERG Potassium Channels Voltage-Gated Neurites Trans-Activators Animals Humans Cation Transport Proteins |
| Zdroj: | Journal of neurobiology. 38(4) |
| ISSN: | 0022-3034 |
| Popis: | The whole-cell patch-clamp technique was used to assess the current carried by inwardly rectifying K+ channels (K(ir)) and the resting membrane potential (RMP) during long-term culture of NG108-15 cells. Culture of this cell line in serum-free medium triggers differentiation of a type I, neuron-like cell type followed by an eventual predominance of a type II, proliferative cell type. NG108-15 K(ir) currents, which strongly resemble currents carried by human ether-a-go-go related gene (HERG) K+ channels, exhibited significantly smaller current density for the more depolarized undifferentiated cells in growth media (GM) and type II cells compared to the neuron-like type I cells. Detailed examination of the transition from undifferentiated GM cells to type I cells revealed a shift in the voltage dependence of K(ir) activation which paralleled the more hyperpolarized RMP, neurite outgrowth, and biochemical differentiation characteristic of type I cells. Reverse-transcription polymerase chain reaction experiments using primers for the rat variant of HERG, RERG, revealed a a nearly twofold increase in RERG mRNA as cells differentiate from GM to type I, a finding entirely consistent with the increased K(ir) current density derived from patch-clamp recordings. Administration of CsCl(5 mM) blocked K(ir) currents and depolarized the RMP of type I cells. Furthermore, culture of NG108-15 cells in serum-free medium but with CsCl added significantly prevented neurite extension, an effect which was entirely reversible upon subsequent removal of CsCl. In contrast, other K+ channel inhibitors (4-aminopyridine and tetraethylammonium), at concentrations without marked effects on K(ir), failed to affect neurite extension. These results suggest an important role of the K(ir) channels in determining the RMP and triggering morphological differentiation of the cell line. |
| Databáze: | OpenAIRE |
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