Nucleotide-induced mucin release from primary hamster tracheal surface epithelial cells involves the P2u purinoceptor.
Mucin release by airway surface epithelial cells is regulated by extracellular adenosine triphosphate (ATP) via a P2 purinoceptor-mediated mechanism. The objective of the present experiment was to examine the possible involvement of uridine triphosphate (UTP) in this purinergic signal transduction pathway. Using primary hamster tracheal surface epithelial cells, ATP and UTP were compared in their abilities: 1) to displace ATP gamma S35-binding to intact cells; 2) to accumulate inositol phosphates; and 3) to stimulate mucin release. Finally, the presence of a P2u receptor message was examined. Our results showed that: 1) UTP was much less effective than ATP in displacing ATP gamma S35-binding (median inhibitory concentrations (IC50S) 240 vs 2.9 microM); 2) UTP was more potent than ATP in accumulating inositol phosphates (100 vs 43% increase at 2mM); 3) UTP was equipotent with ATP in stimulating mucin release; 4) Northern blot analysis of messenger ribonucleic acids (mRNAs) with a mouse P2u receptor complementary deoxyribonucleic acid (cDNA) probe revealed a single specific band (2.8 kb), partial sequencing of which showed a great homology with those of human or mouse P2u receptors. We conclude that, although both ATP and UTP are equipotent in stimulating mucin release, their binding kinetics to the cell surface are quite different, suggesting the presence of a common binding domain which may be responsible for the mucin release by these nucleotides. We suggest that the P2u purinoceptor is likely to be responsible for mucin release by these nucleotides, probably via activation of phospholipase C.
Kim
K C
KC
Dept of Pharmaceutical Sciences, University of Maryland School of Pharmacy, Baltimore, MD 21201, USA.
Park
H R
HR
Shin
C Y
CY
Akiyama
T
T
Ko
K H
KH
D000255:Adenosine Triphosphate / Q000494:pharmacology*; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D002478:Cells, Cultured; D006224:Cricetinae; D004847:Epithelial Cells; D004848:Epithelium / Q000378:metabolism; D008297:Male; D008647:Mesocricetus; D008969:Molecular Sequence Data; D009077:Mucins / Q000187:drug effects / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D012313:RNA / Q000032:analysis; D018048:Receptors, Purinergic P2 / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction; D014132:Trachea / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D014544:Uridine Triphosphate / Q000494:pharmacology*
D009077:Mucins; D018048:Receptors, Purinergic P2; D012313:RNA; D000255:Adenosine Triphosphate; D014544:Uridine Triphosphate