LAN-1: a human neuroblastoma cell line with M1 and M3 muscarinic receptor subtypes coupled to intracellular Ca2+ elevation and lacking Ca2+ channels activated by membrane depolarization

J Neurochem. 1992 Jul;59(1):1-9. doi: 10.1111/j.1471-4159.1992.tb08868.x.


The LAN-1 clone, a cell line derived from a human neuroblastoma, possesses muscarinic receptors. The stimulation of these receptors with increasing concentrations of carbachol (CCh; 1-1,000 microM) caused a dose-dependent increase of the intracellular free Ca2+ concentration ([Ca2+]i). This increase was characterized by an early peak phase (10 s) and a late plateau phase. The removal of extracellular Ca2+ reduced the magnitude of the peak phase to approximately 70% but completely abolished the plateau phase. The muscarinic-activated Ca2+ channel was gadolinium (Gd3+) blockade and nimodipine and omega-conotoxin insensitive. In addition, membrane depolarization did not cause any increase in [Ca2+]i. The CCh-induced [Ca2+]i elevation was concentration-dependently inhibited by pirenzepine and 4-diphenylacetoxy-N-methylpiperidine methiodide, two rather selective antagonists of M1 and M3 muscarinic receptor subtypes, respectively, whereas methoctramine, an M2 antagonist, was ineffective. The coupling of M1 and M3 receptor activation with [Ca2+]i elevation does not seem to be mediated by a pertussis toxin-sensitive guanine nucleotide-binding protein or by the diacylglycerol-protein kinase C system. The mobilization of [Ca2+]i elicited by M1 and M3 muscarinic receptor stimulation seems to be dependent on an inositol trisphosphate-sensitive intracellular store. In addition, ryanodine did not prevent CCh-induced [Ca2+]i mobilization, and, finally, LAN-1 cells appear to lack caffeine-sensitive Ca2+ stores, because the methylxanthine was unable to elicit intracellular Ca2+ mobilization, under basal conditions, after a subthreshold concentration of CCh (0.3 microM), or after thapsigargin.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Calcium / metabolism*
  • Calcium Channel Blockers / pharmacology
  • Calcium Channels / metabolism
  • Calcium Channels / physiology
  • Carbachol / pharmacology
  • Electrophysiology
  • Humans
  • Inositol 1,4,5-Trisphosphate / pharmacology
  • Intracellular Membranes / metabolism
  • Muscarinic Antagonists
  • Neuroblastoma / metabolism*
  • Neuroblastoma / pathology
  • Osmolar Concentration
  • Pertussis Toxin
  • Protein Kinase C / metabolism
  • Receptors, Muscarinic / metabolism*
  • Tumor Cells, Cultured
  • Virulence Factors, Bordetella / pharmacology


  • Calcium Channel Blockers
  • Calcium Channels
  • Muscarinic Antagonists
  • Receptors, Muscarinic
  • Virulence Factors, Bordetella
  • Inositol 1,4,5-Trisphosphate
  • Carbachol
  • Pertussis Toxin
  • Protein Kinase C
  • Calcium