title: Increase in [Ca2+]i by CCh in adult rat sympathetic neurons are not dependent on intracellular Ca2+ pools.
abstract: We have examined the effects of the muscarinic agonists, carbachol (CCh) and oxotremorine (Oxo), on the intracellular free Ca2+ concentration ([Ca2+]i) in acutely dissociated sympathetic neurons from adult rats using fura 2-based microfluorometry. The drugs increased [Ca2+]i by 86 +/- 7 and 38 +/- 10 nM for CCh and Oxo, respectively (both 10 microM). Basal [Ca2+]i was 52 +/- 3 nM. Depletion of the caffeine-sensitive Ca2+ store or blockade of the Ca(2+)-adenosinetriphosphatase with thapsigargin did not alter the effect of either agonist on the rise in [Ca2+]i. On the other hand, the omission of Ca2+ from the perfusion solution or the use of TA-3090, a Ca2+ channel antagonist, blocked the effects of CCh and Oxo. In whole cell current-clamp recordings, the muscarinic agonists elicited a depolarization and action potential firing, which probably explained the rise in [Ca2+]i observed with microfluorimetric recording. In addition to their direct effects on the [Ca2+]i, muscarinic agonists also reduced the rise in [Ca2+]i induced by a nicotinic agonist. This inhibitory effect, observed in 68% of cells that responded to the nicotinic agonist, was blocked by atropine and pertussis toxin, whereas the muscarinic agonist-induced increase in [Ca2+]i was blocked by atropine but was pertussis toxin insensitive. These results suggest that at least two muscarinic receptors are present on sympathetic neurons and that they mediate opposite effect on the fluctuation of [Ca2+]i.
authors.affiliation: Department of Pharmacological and Physiological Science, University of Chicago, Illinois 60637, USA.
authors.forename: S
authors.initials: S
authors.lastname: Foucart
authors.forename: S J
authors.initials: SJ
authors.lastname: Gibbons
authors.forename: J R
authors.initials: JR
authors.lastname: Brorson
authors.forename: R J
authors.initials: RJ
authors.lastname: Miller
mesh_terms: D000818:Animals; D002118:Calcium / Q000378:metabolism*; D002217:Carbachol / Q000494:pharmacology*; D005470:Fluorometry; D007425:Intracellular Membranes / Q000378:metabolism*; D009116:Muscarine / Q000819:agonists; D009474:Neurons / Q000378:metabolism*; D009538:Nicotine / Q000819:agonists; D009994:Osmolar Concentration; D051381:Rats; D013564:Sympathetic Nervous System / Q000166:cytology / Q000378:metabolism*
chemical_list: D009538:Nicotine; D009116:Muscarine; D002217:Carbachol; D002118:Calcium