Ca 2+ mobilization and activation of extracellular acidification by carbachol in acutely dispersed cells from guinea pig detrusor: Fura 2 fluorometry and microphysiometry using the cytosensor

Abstract

The study aim was to develop a simple in vitro model for pharmacophysiological investigation of urinary bladder smooth muscles. Smooth muscle cells from guinea pig detrusor were dissociated, and the suspended cells were stimulated with carbachol (CCh), an acetylcholine receptor agonist. Cytosolic Ca 2+ levels were determined using Fura 2 fluorescence and extracellular acidification rates were monitored by the Cytosensor microphysiometer. CCh dose-dependently increased cytosolic Ca 2+ levels and extracellular acidification rates, with EC 50 values of approximately 1 μM. Both the acetylcholine muscarinic receptor antagonist atropine and the M 3 muscarinic receptor-preferring antagonist 4-diphenylacetoxy N-methylpiperidine (4-DAMP) inhibited the effects of CCh, three orders of magnitude more potently than the selective M 2 muscarinic receptor antagonist, methoctramine. These data indicate the dominant role of M 3 receptors in guinea-pig bladder but fail to show clear evidence of any functional role for M 2 receptors. Since this finding agrees with a number of other studies using in vivo and in vitro models (1), cell suspensions such as these may prove to be simple tools for the pharmacological study of urinary bladder smooth muscle tissue.