| Avtor/Urednik | Benjamin, Elfrida R; Pruthi, Farhana; Olanrewaju, Shakira; Shan, Shen; Hanway, Denise; Liu, Xuesong; Černe, Rok; Lavery, Daniel; Valenzano, Kenneth J; Woodward, Richard M; Ilyin, Victor | |
| Naslov | Pharmacological characterization of recombinant N-type calcium channel (Cav2.2) mediated calcium mobilizytion using FLIPR | |
| Tip | članek | |
| Vir | Biochem Pharmacol | |
| Vol. in št. | Letnik 72 | |
| Leto izdaje | 2006 | |
| Obseg | str. 770-82 | |
| Jezik | eng | |
| Abstrakt | The N-type voltage-gated calcium channel (Cav2.2) functions in neurons to regulate neurotransmitter release. It comprises a clinically relevant target for chronic pain. We have validated a calcium mobilization approach to assessing Cav2.2 pharmacology in two stable Cav2.2 cell lines: alpha1B, alpha2delta, beta3-HEK-293 and alpha1B, beta3-HEK-293. Cav2.2 channels were opened by addition of KCl and Ca2+ mobilization was measured by Fluo-4 fluorescence on a fluorescence imaging plate reader (FLIPR96). Cav2.2 expression and biophysics were confirmed by patchclamp electrophysiology (EP). Both cell lines responded to KCl with adequate signal-to-background. Signals from both cell lines were inhibited by w-conotoxin (ctx)-MVIIa and w-conotoxin (ctx)-GVIa with IC50 values of 1.8 and 1 nM, respectively, for the three-subunit stable, and 0.9 and 0.6 nM, respectively, for the two-subunit stable. Other known Cav2.2 blockers were characterized including cadmium, flunarizine, fluspirilene, and mibefradil. IC50 values correlated with literature EP-derived values. Novel Cav2.2 pharmacology was identified in classes of compounds with other primary pharmacological activities, including Na+ channel inhibitors and antidepressants. Novel Na+ channel compounds with high potency at Cav2.2 were identified in the phenoxyphenyl pyridine, phenoxyphenyl pyrazole, and other classes. The highest potency at Cav2.2 tricyclic antidepressant identified was desipramine. | |
| Deskriptorji | CALCIUM CHANNELS RECOMBINANT PROTEINS ANTIDEPRESSIVE AGENTS SODIUM CHANNELS POTASSIUM CHLORIDE CALCIUM CELL LINE PATCH-CLAMP TECHNIQUES |