Neurotoxic action of veratridine in rat brain neuronal cultures: mechanism of neuroprotection by Ca++ antagonists nonselective for slow Ca++ channels.
The effect of various Ca++ antagonists and local anesthetics on neuronal cell degeneration induced by veratridine was studied in primary rat brain neuronal cultures. Cell death was quantified by measuring lactate dehydrogenase (LDH) released in the culture medium. The neuronal cell degeneration was Ca+(+)-dependent because, in the absence of extracellular Ca++, 16 hr of exposure to 30 microM veratridine failed to produce release of LDH. Ca++ antagonists, nonselective for slow Ca++ channels (flunarizine, cinnarizine, lidoflazine, prenylamine and bepridil) inhibited veratridine-induced release of LDH with IC50 values between 0.11 and 0.47 microM. Ca++ antagonists selective for slow Ca++ channels were less potent and inhibited veratridine-induced release of LDH at concentrations in the following order of potency: nicardipine greater than gallopamil and verapamil greater than niludipine greater than nitrendipine greater than nifedipine greater than nimodipine greater than diltiazem. Tested local anesthetics were incomplete inhibitors of veratridine-induced release of LDH. A good correlation was found between the potency of the drugs to inhibit released LDH induced by 30 microM veratridine in neuronal cultures and their binding affinity for the batrachotoxin binding site of Na+ channels in rat cortex synaptosomal preparation. It is concluded that protection against veratridine-induced neurotoxicity can be mediated by blocking a veratridine-sensitive Na+ channel. It is a property of certain nonselective Ca++ antagonists. There is apparently no direct relationship with Ca++ antagonistic activity. The effect is unrelated to local anesthetic activity.
Pauwels
P J
PJ
Department of Biochemical Pharmacology, Janssen Research Foundation, Beerse, Belgium.
Van Assouw
H P
HP
Peeters
L
L
Leysen
J E
JE
D000777:Anesthetics / Q000494:pharmacology; D000818:Animals; D001496:Batrachotoxins / Q000378:metabolism; D001665:Binding Sites; D001921:Brain / Q000166:cytology / Q000187:drug effects / Q000201:enzymology / Q000648:ultrastructure; D001927:Brain Diseases / Q000139:chemically induced / Q000201:enzymology / Q000517:prevention & control; D002121:Calcium Channel Blockers / Q000494:pharmacology; D015220:Calcium Channels / Q000378:metabolism; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D003600:Cytosol / Q000201:enzymology; D005260:Female; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D009474:Neurons / Q000166:cytology / Q000187:drug effects* / Q000201:enzymology; D051381:Rats; D011919:Rats, Inbred Strains; D015222:Sodium Channels / Q000378:metabolism; D013574:Synaptosomes / Q000378:metabolism / Q000648:ultrastructure; D014316:Tritium; D014701:Veratridine / Q000633:toxicity*
D000777:Anesthetics; D001496:Batrachotoxins; D002121:Calcium Channel Blockers; D015220:Calcium Channels; D015222:Sodium Channels; D014316:Tritium; D014701:Veratridine; C031775:batrachotoxinin A 20-alpha-benzoate; D007770:L-Lactate Dehydrogenase