Distinct effects of omega-toxins and various groups of Ca(2+)-entry inhibitors on nicotinic acetylcholine receptor and Ca2+ channels of chromaffin cells

Eur J Pharmacol. 1997 Feb 12;320(2-3):249-57. doi: 10.1016/s0014-2999(96)00902-8.

Abstract

The effects of omega-toxins and various Ca2+ antagonist subtypes on the 45Ca2+ entry into bovine adrenal medullary chromaffin cells stimulated via nicotinic acetylcholine receptors or via direct depolarization with K+, have been compared. The conditions selected to stimulate the 45Ca2+ entry consisted of a 60-s period of exposure of cells to 100 microM of the nicotinic acetylcholine receptor agonist dimethylphenylpiperazinium or to 70 mM K+. The N-type voltage-dependent Ca2+ channel blockers omega-conotoxin GVIA and MVIIA (1 microM) inhibited 45Ca2+ entry stimulated by dimethylphenylpiperazinium or K+ by around 25-30%. The P-type Ca2+ channel blocker omega-agatoxin IVA (10 nM) did not affect the dimethylphenylpiperazinium nor the K+ responses; 1 microM (Q-channel blockade) inhibited both responses by around 50%. The N/P/Q-type Ca2+ channel blocker omega-contoxin MVIIC (1 microM) inhibited the K+ evoked 45Ca2+ entry by 70%, while dimethylphenylpiperazinium was blocked by 50% (P < 0.001). The L-type Ca2+ channel blockers nifedipine, furnidipine, diltiazem or verapamil (3 microM each) inhibited much more the dimethylphenylpiperazinium than the K+ response. The dimethylphenylpiperazinium signal was blocked 71, 88, 89, and 53%, respectively, by nifedipine, furnidipine, diltiazem and verapamil, and the K+ response by 38, 29, 22, and 10%. Combined omega-conotoxin MVIIC (1 microM) and furnidipine (3 microM) blocked 100% of the K+ evoked 45Ca2+ entry. However, combined omega-conotoxin GVIA (1 microM), and furnidipine left unblocked 50% of the K+ response. The "wide spectrum' Ca2+ channel antagonists flunarizine or dotarizine (3 microM each) blocked the dimethylphenylpiperazinium and the K+ responses to a similar extent (50%); cinnarizine (3 microM) inhibited more the dimethylphenylpiperazinium (82%) than the K+ response (21%). At 3 microM, the highly lipophilic beta-adrenoceptor antagonist (+/-)-propranolol, reduced by 68% the dimethylphenylpiperazinium signal and by 23% the K+ signal. Other high lipophilic beta-adrenoceptor antagonists such as metoprolol and labetalol, reduced little the dimethylphenylpiperazinium and the K+ responses. The highly lipophilic agent penfluridol blocked the dimethylpiperazinium response by 30% and the K+ response by 50%. One of the least lipophilic compounds tested, (+)-lubeluzole, blocked by 40% the dimethylphenylpiperazinium and the K+ responses. These data are compatible with the idea that the various omega-toxin peptides used to separate pharmacologically the different voltage-dependent Ca2+ channels expressed by neurones, do not block the neuronal nicotinic acetylcholine receptor ion channel. In contrast the L-type Ca2+ channel blockers do block the nicotinic acetylcholine receptor ionophore. Lipophilicity of the compounds is not a requirement for Ca2+ channel or nicotinic acetylcholine receptor blockade.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Calcium / pharmacokinetics
  • Calcium Channel Blockers / pharmacology*
  • Calcium Channels / drug effects*
  • Calcium Channels / metabolism
  • Calcium Radioisotopes
  • Cattle
  • Chromaffin Cells / drug effects*
  • Chromaffin Cells / metabolism
  • Chromaffin Cells / ultrastructure*
  • Dimethylphenylpiperazinium Iodide
  • Peptides / pharmacology*
  • Potassium / pharmacology
  • Receptors, Nicotinic / drug effects*
  • Spider Venoms / pharmacology*
  • Stimulation, Chemical
  • omega-Agatoxin IVA
  • omega-Conotoxin GVIA
  • omega-Conotoxins*

Substances

  • Calcium Channel Blockers
  • Calcium Channels
  • Calcium Radioisotopes
  • Peptides
  • Receptors, Nicotinic
  • Spider Venoms
  • omega-Agatoxin IVA
  • omega-Conotoxins
  • Dimethylphenylpiperazinium Iodide
  • ziconotide
  • omega-Conotoxin GVIA
  • Potassium
  • Calcium