Endogenous ganglioside GM1 modulates L-type calcium channel activity in N18 neuroblastoma cells
| Autor: | Daniel H. Mascó, Gary Brooker, R. O. Carlson, Sarah Spiegel |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 1994 |
| Předmět: |
Cholera Toxin
Dihydropyridines P-type calcium channel Fura-2 chemistry.chemical_element G(M1) Ganglioside N-type calcium channel Calcium Cell Line chemistry.chemical_compound Mice Neuroblastoma Nicardipine Nickel Tumor Cells Cultured Animals G(M3) Ganglioside L-type calcium channel Egtazic Acid Manganese Chemistry General Neuroscience Calcium channel T-type calcium channel Articles 3-Pyridinecarboxylic acid 1 4-dihydro-2 6-dimethyl-5-nitro-4-(2-(trifluoromethyl)phenyl)- Methyl ester Calcium Channel Blockers R-type calcium channel Biochemistry Microscopy Fluorescence Biophysics Potassium Calcium Channels |
| Popis: | Digital imaging fluorescence microscopy was used to investigate the effect of the B subunit of cholera toxin on calcium homeostasis in neuroblastoma N18 cells. The B subunit, which binds specifically to ganglioside GM1 in the outer leaflet of the cell membrane, was found to induce a sustained increase of intracellular calcium concentration ([Ca2+]i). The increase in [Ca2+]i was not observed in the absence of extracellular calcium, or in the presence of the calcium chelator EGTA, and was blocked by nickel. The B subunit was also found to induce an influx of manganese ions, as indicated by a quench of the intracellular fura-2 fluorescence. These data suggest that the B subunit induces an increase in calcium influx in N18 cells. Potassium-induced depolarization also stimulated manganese influx; however, after the onset of depolarization-induced influx, the B subunit had no further effect. This occlusion suggests involvement of voltage-dependent calcium channels. Treatment with BayK8644, a dihydropyridine agonist selective for L-type calcium channels, induced manganese influx that was not altered by the B subunit and apparently blocked the effect of the B subunit itself. Furthermore, the dihydropyridine L-type channel antagonists niguldipine or nicardipine completely inhibited B subunit- induced manganese influx. Thus, the B subunit-induced manganese influx is likely due to activation of an L-type voltage-dependent calcium channel. Spontaneous influx of manganese ions was also inhibited by nicardipine or niguldipine and by exogenous gangliosides. Ganglioside GM1 was more potent than GM3, but globoside had no significant effect. The modulation of L-type calcium channels by endogenous ganglioside GM1 has important implications for its role in neural development, differentiation, and regeneration and also for its potential function in the electrical excitability of neurons. |
| Databáze: | OpenAIRE |
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