Polarized distribution of bradykinin receptors on airway epithelial cells and independent coupling to second messenger pathways.
Bradykinin stimulates Cl- secretion by airway epithelia, but different patterns of secretion result from addition to the mucosal and submucosal surfaces. Earlier work suggested that bradykinin activates two second messenger pathways: increasing inositol phosphates (InsP) via phosphatidylinositol bisphosphate hydrolysis and increasing cAMP via arachidonic acid metabolism. In this study, we measured arachidonic acid release and InsP production in cultured canine tracheal epithelial cells. Bradykinin increased the two second messengers via independent mechanisms: (a) dose-response curves with different incubation media demonstrated that each second messenger could be generated independently of the other; (b) phorbol ester inhibited InsP production but stimulated arachidonic acid release; (c) for polarized cultures, submucosal bradykinin stimulated production of both second messengers but mucosal bradykinin stimulated only arachidonic acid release. To determine if differences in second messenger formation at the two membranes resulted from differences in hormone-receptor interactions, we compared bradykinin binding to the apical and basolateral membranes. Both the binding capacities and affinity constants (KD) were different (basolateral KD, 257 +/- 53 pM; apical KD, 39 +/- 3 pM). These data demonstrate polarized coupling of bradykinin receptors to second messenger pathways in airway epithelial cells and suggest that this polarized coupling is due to different bradykinin receptors at the two membranes.
Denning
G M
GM
Howard Hughes Medical Institute, Department of Internal Medicine, University of Iowa College of Medicine, Iowa City 52242.
Welsh
M J
MJ
D000818:Animals; D016718:Arachidonic Acid; D001095:Arachidonic Acids / Q000378:metabolism*; D001920:Bradykinin / Q000378:metabolism* / Q000494:pharmacology; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D002478:Cells, Cultured; D004285:Dogs; D004848:Epithelium / Q000187:drug effects / Q000378:metabolism; D016244:Guanosine 5'-O-(3-Thiotriphosphate) / Q000494:pharmacology; D007295:Inositol Phosphates / Q000378:metabolism*; D007700:Kinetics; D018002:Receptors, Bradykinin; D017981:Receptors, Neurotransmitter / Q000187:drug effects / Q000378:metabolism / Q000502:physiology*; D015290:Second Messenger Systems / Q000187:drug effects; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014132:Trachea / Q000378:metabolism*
D001095:Arachidonic Acids; D007295:Inositol Phosphates; D018002:Receptors, Bradykinin; D017981:Receptors, Neurotransmitter; D016718:Arachidonic Acid; D016244:Guanosine 5'-O-(3-Thiotriphosphate); D013755:Tetradecanoylphorbol Acetate; D001920:Bradykinin