Characterisation of the L- and N-type calcium channels in differentiated SH-SY5Y neuroblastoma cells: calcium imaging and single channel recording

Brain Res Mol Brain Res. 1992 Mar;13(1-2):53-61. doi: 10.1016/0169-328x(92)90044-c.

Abstract

We have used single cell imaging of [Ca2+]i and single channel cell-attached patch clamp recording to characterise the Ca2+ channels present on the plasma membrane of retinoic acid-differentiated human neuroblastoma (SH-SY5Y) cells. Exposure to raised K+ (45 or 60 mM) for 1 min resulted in a transient rise in [Ca2+]i which was abolished by cadmium (100 microM). The amplitude of the evoked rise varied from cell to cell. Both omega-Conus toxin (500 nM) and nifedipine (10 microM) reduced, but did not abolish, the rise in [Ca2+]i whereas Bay K 8644 (3 microM) potentiated it. In single channel records both L- and N-type Ca2+ channel openings were observed during membrane depolarisations from a holding potential of -90 mV. L-type channel openings (unitary conductance 22.5 pS) were prolonged by S(+)-PN 202-791 (500 nM) and could still be evoked from a depolarised holding potential (-40 mV). N-type channel openings (unitary conductance 12.5 pS) were unaffected by the dihydropyridine agonist but were inactivated at a holding potential of -40 mV. These results indicate that, in contrast to previous observations using whole cell recording, retinoic acid-differentiated SH-SY5Y cells express both L- and N-type Ca2+ channels.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethyl-5-nitro-4-(2-(trifluoromethyl)phenyl)-, Methyl ester / pharmacology
  • Cadmium / pharmacology
  • Cadmium Chloride
  • Calcium / metabolism*
  • Calcium Channel Blockers / pharmacology*
  • Calcium Channels / drug effects
  • Calcium Channels / physiology*
  • Calcium Channels / ultrastructure
  • Cell Differentiation / drug effects
  • Cell Line
  • Cell Membrane / metabolism
  • Chlorides / pharmacology
  • Evoked Potentials / drug effects
  • Fura-2
  • Humans
  • Kinetics
  • Microscopy, Fluorescence
  • Neuroblastoma
  • Nicotinic Acids / pharmacology
  • Nifedipine / pharmacology
  • Oxadiazoles*
  • Peptides, Cyclic / pharmacology
  • Potassium / pharmacology
  • Tretinoin / pharmacology
  • omega-Conotoxins*

Substances

  • Calcium Channel Blockers
  • Calcium Channels
  • Chlorides
  • Nicotinic Acids
  • Oxadiazoles
  • Peptides, Cyclic
  • omega-Conotoxins
  • Cadmium
  • PN 202-791
  • Conus magus toxin
  • Tretinoin
  • 3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethyl-5-nitro-4-(2-(trifluoromethyl)phenyl)-, Methyl ester
  • Nifedipine
  • Cadmium Chloride
  • Potassium
  • Calcium
  • Fura-2