title: Characterization of Ca2+ signaling pathways in mouse adrenal medullary chromaffin cells.
abstract: In the present study, we characterized the Ca2+ responses and secretions induced by various secretagogues in mouse chromaffin cells. Activation of the acetylcholine receptor (AChR) by carbachol induced a transient intracellular Ca2+ concentration ([Ca2+](i)) increase followed by two phases of [Ca2+](i) decay and a burst of exocytic events. The contribution of the subtypes of AChRs to carbachol-induced responses was examined. Based on the results obtained by stimulating the cells with the nicotinic receptor (nAChR) agonist, 1,1-dimethyl-4-phenylpiperazinium iodide, high K(+) and the effects of thapsigargin, it appears that activation of nAChRs induces an extracellular Ca2+ influx, which in turn activate Ca(2+)-induced Ca2+ release via the ryanodine receptors. Muscarine, a muscarinic receptor (mAChRs) agonist, was found to induce [Ca2+](i) oscillation and sustained catecholamine release, possibly by activation of both the receptor- and store-operated Ca2+ entry pathways. The RT-PCR results showed that mouse chromaffin cells are equipped with messages for multiple subtypes of AChRs, ryanodine receptors and all known components of the receptor- and store-operated Ca2+ entry. Furthermore, results obtained by directly monitoring endoplasmic reticulum (ER) and mitochondrial Ca2+ concentration and by disabling mitochondrial Ca2+ uptake suggest that the ER acts as a Ca2+ source, while the mitochondria acts as a Ca2+ sink. Our results show that both nAChRs and mAChRs contribute to the initial carbachol-induced [Ca2+](i) increase which is further enhanced by the Ca2+ released from the ER mediated by Ca(2+)-induced Ca2+ release and mAChR activation. This information on the Ca2+ signaling pathways should lay a good foundation for future studies using mouse chromaffin cells as a model system.
authors.affiliation: Molecular Medicine Program, National Yang-Ming University, Taipei, Taiwan.
authors.forename: Pei-Chun
authors.initials: PC
authors.lastname: Wu
authors.forename: Ming-Ji
authors.initials: MJ
authors.lastname: Fann
authors.forename: Lung-Sen
authors.initials: LS
authors.lastname: Kao
mesh_terms: D000313:Adrenal Medulla / Q000166:cytology*; D000818:Animals; D002110:Caffeine / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism*; D020013:Calcium Signaling / Q000187:drug effects / Q000502:physiology*; D002217:Carbachol / Q000494:pharmacology; D002395:Catecholamines / Q000378:metabolism; D018678:Cholinergic Agents / Q000494:pharmacology; D019439:Chromaffin Cells / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D004246:Dimethylphenylpiperazinium Iodide / Q000494:pharmacology; D004347:Drug Interactions; D004721:Endoplasmic Reticulum / Q000187:drug effects / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005089:Exocytosis / Q000187:drug effects; D051379:Mice; D008810:Mice, Inbred C57BL; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism; D009116:Muscarine / Q000494:pharmacology; D010726:Phosphodiesterase Inhibitors / Q000494:pharmacology; D011189:Potassium Chloride / Q000494:pharmacology; D012333:RNA, Messenger / Q000378:metabolism; D011950:Receptors, Cholinergic / Q000235:genetics / Q000378:metabolism; D019284:Thapsigargin / Q000494:pharmacology; D013997:Time Factors
chemical_list: D002395:Catecholamines; D018678:Cholinergic Agents; D004791:Enzyme Inhibitors; D010726:Phosphodiesterase Inhibitors; D012333:RNA, Messenger; D011950:Receptors, Cholinergic; D002110:Caffeine; D004246:Dimethylphenylpiperazinium Iodide; D011189:Potassium Chloride; D019284:Thapsigargin; D009116:Muscarine; D002217:Carbachol; D002118:Calcium