oxonol-v and Neuroblastoma

Membrane potential was measured in the suspension of SH-SY5Y cells using the anionic potentiometric probe, oxonol V. The relation of fluorescence to membrane potential was assessed by increasing the external [K+] in the presence of the K+ ionophore valinomycin. The response was linear in the range of 5 to 30 mM K+ (membrane potential change of approximately 40 mV). Muscarine increased the fluorescence indicating a depolarization. The competitive inhibitory constant (112 nM) of the muscarinic antagonist pirenzepine (5,11-dihydro-11-([4-methyl-1-piperazinyl]acetyl)-6H-pyrido[2,3-b] (1,4)benzodiazepin-6-one-dihydrochloride) suggests that Hm1 receptors are not involved. The protein kinase C inhibitor, GF 109203X (3-[1-(3-demethylaminopropyl)-indol-3-yl]-3-(indol-3-yl)-maleimide ), and a reduction of extracellular Na+ both produced an additive partial inhibition. The results suggest that muscarinic receptors depolarize these cells by separate Na(+)-dependent and -independent mechanisms, the Na(+)-independent mechanism being protein kinase C-dependent.

Topics: Egtazic Acid; Electrophysiology; Fluorescent Dyes; Humans; Indicators and Reagents; Isoxazoles; Membrane Potentials; Muscarine; Muscarinic Agonists; Neuroblastoma; Receptors, Muscarinic; Tetrodotoxin; Time Factors; Tumor Cells, Cultured