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Journal of Cell Science, Vol 107, Issue 10 2899-2907, Copyright © 1994 by Company of Biologists

JOURNAL ARTICLES

cAMP does not regulate [Ca2+]i in human tracheal epithelial cells in primary culture

PB Davis, CL Silski and A Perez
Department of Pediatrics, Case Western Reserve University at Rainbow Babies and Childrens Hospital, Cleveland, Ohio 44106.

Human tracheal epithelial cells in primary culture respond to different receptor agonists with different peak intracellular calcium concentrations. From resting concentration 138 +/- 13 nM, bradykinin (0.1 microM) produces an increase to a maximum of 835 +/- 195 nM, histamine (10 microM) to 352 +/- 51 nM, and ATP (5-500 microM) to more than 1500 nM. Nine of 14 cultures also responded to isoproterenol (10 microM), though with a smaller increase, to 210 +/- 29 nM. A response was observed with isoproterenol, and epinephrine, but not norepinephrine, phenylephrine or methoxamine, was inhibited by propranolol but not phentolamine, and so this appeared to be a beta-adrenergic response. However, no response could be detected to adenosine, prostaglandin E2 or forskolin, agents that activate adenylate cyclase, or to permeant analogs of cAMP (CPT-cAMP or db-cAMP). The intracellular calcium response to isoproterenol did not follow either the time-course or the desensitization pattern of the cAMP response. Thus, this response to isoproterenol is not mediated by cAMP. No relation was demonstrated between cAMP production by other agonists and the response of intracellular calcium. Pretreatment with agents that increase cAMP did not affect the calcium responses to ATP or bradykinin. Thus, cAMP does not regulate intracellular calcium concentration in human tracheal epithelial cells. The variation in peak intracellular calcium responses to various agonists may be explained by the presence of multiple second messengers (other than cAMP), multiple intracellular pools of calcium, or cell heterogeneity. The agonists tested had the same relative potency in cells from patients with cystic fibrosis as in non-cystic fibrosis cells.


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© The Company of Biologists Ltd 1994