title: Cholinergic stimulation enhances cytosolic calcium ion accumulation in mouse hippocampal CA1 pyramidal neurones during short action potential trains.
abstract: Acetylcholine is a regulatory cofactor for numerous activity-dependent processes of central nervous system development and plasticity in which increases in cytosolic calcium ion concentration ([Ca(2+)](cyto) couple membrane excitation to cellular changes. We examined how cholinergic receptor activation affects temporal and spatial aspects of increases in [Ca(2+)](cyto) during short trains of action potentials in hippocampal CA1 pyramidal neurones. Membrane-impermeant Ca(2+)-sensitive dye was introduced into the cytosol during whole-cell recordings, and Ca(2+)-dependent fluorescence was recorded from somatic, nuclear and proximal dendrite regions with high temporal resolution. In all neuronal compartments, the cholinergic agonist carbachol (5 microM) increased resting [Ca(2+)](cyto) and the maximum [Ca(2+)](cyto) attained during a short action potential train. Carbachol also slowed the recovery of [Ca(2+)](cyto) towards resting levels. The largest increases in peak cytosolic Ca(2+) concentration (delta [Ca(2+)](cyto) were seen in the dendrite and apical cell body, while relaxations of the carbachol-induced increase in delta [Ca(2+)](cyto) showed greater prolongation in the nucleus and basal cell body. Most significantly, the difference between Ca(2+) signals recorded before and during exposure to carbachol consistently showed a monotonic rise and smooth fall in all cell compartments, suggesting that the increase in [Ca(2+)](cyto) associated with each action potential was not altered by carbachol. Consistent with this view, changes in Ca(2+) signalling were not accompanied by changes in action potential waveforms. The effects of carbachol were partially reversed by simultaneous exposure to atropine, or partially inhibited by inclusion of heparin in the intracellular solution, indicating the involvement of muscarinic acetylcholine receptors and InsP(3)-sensitive Ca(2+)-release channels. Our data indicate that carbachol-induced slowing of [Ca(2+)]cyto relaxations after each action potential results in enhanced accumulation of Ca(2+) in the cytosol in the absence of changes in action potential-driven Ca(2+) entry. By modulating the time course of Ca(2+) signals, cholinergic stimulation may regulate the activation of Ca(2+)-dependent intracellular processes dependent on patterns of [Ca(2+)](cyto) changes.
authors.affiliation: Division of Neurosciences, Beckman Research Institute of the City of Hope, Duarte, CA 91010, USA.
authors.forename: S M
authors.initials: SM
authors.lastname: Beier
authors.forename: M E
authors.initials: ME
authors.lastname: Barish
mesh_terms: D000200:Action Potentials / Q000187:drug effects; D000818:Animals; D000831:Animals, Newborn; D001285:Atropine / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism*; D002217:Carbachol / Q000494:pharmacology; D002451:Cell Compartmentation / Q000187:drug effects / Q000502:physiology; D018678:Cholinergic Agents / Q000494:pharmacology*; D018679:Cholinergic Agonists / Q000494:pharmacology; D003600:Cytosol / Q000187:drug effects / Q000378:metabolism*; D004558:Electric Stimulation; D005456:Fluorescent Dyes; D006493:Heparin / Q000494:pharmacology; D006624:Hippocampus / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D066298:In Vitro Techniques; D051379:Mice; D018727:Muscarinic Antagonists / Q000494:pharmacology; D017966:Pyramidal Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D011976:Receptors, Muscarinic / Q000187:drug effects / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects
chemical_list: D018678:Cholinergic Agents; D018679:Cholinergic Agonists; D005456:Fluorescent Dyes; D018727:Muscarinic Antagonists; D011976:Receptors, Muscarinic; D001285:Atropine; D002217:Carbachol; D006493:Heparin; D002118:Calcium