title: Potent stimulation of large-conductance Ca2+-activated K+ channels by rottlerin, an inhibitor of protein kinase C-delta, in pituitary tumor (GH3) cells and in cortical neuronal (HCN-1A) cells.
abstract: The effects of rottlerin, a known inhibitor of protein kinase C-delta activation, on ion currents were investigated in pituitary tumor (GH3) cells. Rottlerin (0.3-100 microM) increased the amplitude of Ca2+-activated K+ current (I K(Ca)) in a concentration-dependent manner with an EC50 value of 1.7 microM. In intracellular perfusion with rottlerin (1 microM) or staurosporine (10 microM), phorbol 12-myristate 13-acetate-induced inhibition of I K(Ca) in these cells was abolished. In cell-attached mode, rottlerin applied on the extracellular side of the membrane caused activation of large-conductance Ca2+-activated K+ (BK(Ca)) channels, and a further application of BAPTA-AM (10 microM) to the bath had no effect on rottlerin-stimulated channel activity. When cells were exposed to rottlerin, the activation curve of these channels was shifted to less positive potential with no change in the slope factor. Rottlerin increased BK(Ca)-channel activity in outside-out patches. Its change in kinetic behavior of BK(Ca) channels is primarily due to an increase in mean open time. With the aid of minimal kinetic scheme, a quantitative description of rottlerin stimulation on BK(Ca) channels in GH3 cells was also provided. Under current-clamp configuration, rottlerin (1 microM) decreased the firing of action potentials. I K(Ca) elicited by simulated action potential waveforms was enhanced by this compound. In human cortical HCN-1A cells, rottlerin (1 microM) could also interact with the BK(Ca) channel to stimulate I K(Ca). Therefore, rottlerin may directly activate BK(Ca) channels in neurons or endocrine cells.
authors.affiliation: Department of Physiology, National Cheng Kung University Medical College, Tainan, Taiwan. snwu@mail.ncku.edu.tw
authors.forename: Sheng-Nan
authors.initials: SN
authors.lastname: Wu
authors.forename: Ya-Jean
authors.initials: YJ
authors.lastname: Wang
authors.forename: Ming-Wei
authors.initials: MW
authors.lastname: Lin
mesh_terms: D000098:Acetophenones / Q000494:pharmacology*; D000200:Action Potentials / Q000187:drug effects; D000236:Adenoma / Q000378:metabolism* / Q000473:pathology; D000818:Animals; D001578:Benzopyrans / Q000494:pharmacology*; D002273:Carcinogens / Q000494:pharmacology; D002460:Cell Line; D045744:Cell Line, Tumor; D002540:Cerebral Cortex / Q000166:cytology / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D006801:Humans; D051037:Large-Conductance Calcium-Activated Potassium Channel alpha Subunits; D009474:Neurons / Q000166:cytology / Q000378:metabolism*; D010911:Pituitary Neoplasms / Q000378:metabolism* / Q000473:pathology; D015221:Potassium Channels / Q000187:drug effects / Q000378:metabolism; D024681:Potassium Channels, Calcium-Activated / Q000187:drug effects* / Q000378:metabolism; D051745:Protein Kinase C-delta / Q000037:antagonists & inhibitors* / Q000502:physiology; D051381:Rats; D019311:Staurosporine / Q000494:pharmacology; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
chemical_list: D000098:Acetophenones; D001578:Benzopyrans; D002273:Carcinogens; D004791:Enzyme Inhibitors; C494842:Kcnma1 protein, rat; D051037:Large-Conductance Calcium-Activated Potassium Channel alpha Subunits; D015221:Potassium Channels; D024681:Potassium Channels, Calcium-Activated; C085746:rottlerin; D051745:Protein Kinase C-delta; D019311:Staurosporine; D013755:Tetradecanoylphorbol Acetate