Maitotoxin induces a calcium-dependent membrane depolarization in GH4C1 pituitary cells via activation of Type L voltage-dependent calcium channels

• Published in: The Journal of biological chemistry 1992-12, Vol.267 (35), p.25025-25031
• Main Authors: DAN XI, VAN DOLAH, F. M, RAMSDELL, J. S
• Format: Article
• Language: English
• Subjects: algae; Animals; Bacteriology; Biological and medical sciences; calcio; calcium; Calcium - metabolism; Calcium Channels - drug effects; Calcium Channels - physiology; cation; cationes; cations; cell culture; Cell Membrane - drug effects; Cell Membrane - physiology; Choline - pharmacology; cultivo de celulas; culture de cellule; Dose-Response Relationship, Drug; envenenamiento; Fundamental and applied biological sciences. Psychology; intoxication; Isradipine - metabolism; Kinetics; Marine Toxins - pharmacology; membrana; membrane; Membrane Potentials - drug effects; membranes; Microbiology; Nimodipine - pharmacology; Oxocins; Pathogenicity, virulence, toxins, bacteriocins, pyrogens, host-bacteria relations, miscellaneous strains; Pituitary Neoplasms; poisoning; Potassium - pharmacology; Rats; Sodium - pharmacology; Tumor Cells, Cultured
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1016/S0021-9258(19)74000-9

Maitotoxin (MTX) is a water-soluble polyether, isolated from the marine dinoflagellate Gambierdiscus toxicus, that stimulates hormone release and Ca(2+) influx. We have investigated the action by which MTX induces Ca(2+) influx and stimulates prolactin (PRL) release from GH4C1 rat pituitary cells. PRL release elicited by MTX is abolished in a concentration-dependent manner by nimodipine, a dihydropyridine (DHP) antagonist of type L voltage-dependent calcium channels (L-VDCC), indicating that MTX-enhanced PRL release occurs via activation of type L-VDCC. As an initial approach to determine whether MTX interacts directly with VDCC, we examined whether MTX affects the binding of [3H]PN 200-110, a DHP class antagonist, in intact GH4C1 cells. MTX increased the Bmax of [3H]PN 200-110 binding to intact GH4C1 cells from 4.6 +/- 0.03 to 12.5 +/- 2.2 fmol/10(6) cells, without changing the Kd. This indicates that MTX does not bind to the DHP site, but rather suggests that MTX may have an allosteric interaction with the DHP binding site. The effect of MTX on DHP binding was largely (65%) calcium-dependent. We next examined whether MTX alters the membrane potential of GH4C1 cells using the potential sensitive fluorescent dye bisoxonol. Addition of 100 ng/ml MTX to GH4C1 cells caused a membrane depolarization within 2.5 min which reached a plateau at 5 min. The MTX-induced depolarization was not prevented by substitution of impermeant choline ions for Na+. It was similarly unaffected by K+ channel blockers or by depleting the K+ chemical concentration gradient with gramicidin, a monovalent cation pore-forming agent. By contrast, low extracellular Ca(2+) totally abolished the depolarization response, and nimodipine at 100 nM substantially reduced the MTX-induced membrane depolarization. These results indicate that the predominant effect of MTX on depolarization is Ca(2+) influx through L-VDCC. Taken together, our results indicate that MTX-enhanced PRL release occurs exclusively via activation of type L-VDCC in GH4C1 cells. We suggest that MTX induces an initial slow calcium conductance, possibly via an allosteric interaction with a component of the VDCC complex, which, in turn, initiates a positive feedback mechanism involving calcium-dependent membrane depolarization and voltage-dependent activation of calcium channels.