UDP activates a mucosal-restricted receptor on human nasal epithelial cells that is distinct from the P2Y2 receptor.
The presence of the P2Y2 (P(2U)-purinergic) receptor on the apical surface of airway tissue raises the possibility that aerosolized UTP might be used therapeutically to induce Cl- secretion in individuals with cystic fibrosis. However, the duration of the effects of UTP may be limited by enzymatic degradation. We therefore have analyzed the metabolism of UTP and its metabolite UDP on polarized human nasal epithelium (HNE), and have compared the pharmacological activities of these two uridine nucleotides. HPLC analysis of medium bathing the mucosal surface of HNE cells revealed the presence of an ecto-nucleotidase(s) that hydrolyzed [3H]UTP and [3H]UDP with t1/2 values (at 1 microM nucleotide) of 14 and 27 min, respectively. An ecto-nucleoside diphosphokinase activity also was observed, which promoted conversion of [3H]UDP into [3H]UTP in the presence of ATP. The effects of UDP on [3H]inositol phosphate accumulation, intracellular calcium levels ([Ca2+]i), and Cl- secretory rates (I(Cl-)) were quantitated in HNE cells in the presence of hexokinase and glucose to ensure that no UTP (or ATP) contaminated UDP solutions during the assays. Although UDP does not activate the human P2Y2 receptor, mucosal addition of UDP promoted [3H]inositol phosphate accumulation with an EC50 of 190 nM. Mucosal addition of UTP stimulated [3H]inositol phosphate accumulation with an EC50 of 280 nM. The maximal effects of mucosal UDP on [3H]inositol phosphate, [Ca2+]i, and I(Cl-) responses were approximately one-half of those observed with mucosal UTP. Serosal application of UTP promoted a 50% greater [3H]inositol phosphate and calcium response than did mucosal application of UTP. In contrast, UDP had no effect when added to the serosal medium. Repetitive mucosal applications of UDP to HNE cells resulted in a progressive loss, i.e., desensitization, of the [Ca2+]i and I(Cl-) response to UDP, whereas the corresponding responses to UTP remained unchanged. Our results provide evidence for the existence of a UDP receptor on HNE cells that is pharmacologically distinct from the P2Y2 receptor. The relative stability of UDP on the airway surface and the apparent predominant mucosal expression of this putative UDP receptor make it a potential target for cystic fibrosis treatment.
Lazarowski
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ER
Department of Pharmacology, School of Medicine, University of North Carolina, Chapel Hill 27599, USA.
Paradiso
A M
AM
Watt
W C
WC
Harden
T K
TK
Boucher
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RC
D002118:Calcium / Q000378:metabolism; D002478:Cells, Cultured; D002712:Chlorides / Q000378:metabolism; D006593:Hexokinase / Q000378:metabolism; D006801:Humans; D007295:Inositol Phosphates / Q000378:metabolism; D007700:Kinetics; D009297:Nasal Mucosa / Q000187:drug effects / Q000502:physiology*; D009701:Nucleoside-Diphosphate Kinase / Q000378:metabolism; D011956:Receptors, Cell Surface / Q000187:drug effects / Q000502:physiology*; D018048:Receptors, Purinergic P2 / Q000502:physiology*; D058473:Receptors, Purinergic P2Y2; D014420:Turbinates; D014530:Uridine Diphosphate / Q000378:metabolism / Q000494:pharmacology*; D014544:Uridine Triphosphate / Q000378:metabolism / Q000494:pharmacology*
D002712:Chlorides; D007295:Inositol Phosphates; C546385:P2RY2 protein, human; D011956:Receptors, Cell Surface; D018048:Receptors, Purinergic P2; D058473:Receptors, Purinergic P2Y2; D014530:Uridine Diphosphate; D006593:Hexokinase; D009701:Nucleoside-Diphosphate Kinase; D002118:Calcium; D014544:Uridine Triphosphate