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Am J Physiol Lung Cell Mol Physiol 280: L98-L106, 2001;
1040-0605/01 $5.00
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Vol. 280, Issue 1, L98-L106, January 2001

Extracellular cyclic ADP-ribose potentiates ACh-induced contraction in bovine tracheal smooth muscle

Luisa Franco1, Santina Bruzzone2, Pinfang Song3, Lucrezia Guida2, Elena Zocchi2, Timothy F. Walseth4, Emanuele Crimi3, Cesare Usai5, Antonio De Flora2, and Vito Brusasco3

1 Biocrystallography Centre-Consiglio Nazionale delle Ricerche, University Federico II, 80134 Naples; 2 Section of Biochemistry, Department of Experimental Medicine, and 3 Department of Motor Sciences, University of Genova, 16132 Genoa; 5 Institute of Cybernetics and Biophysics, National Research Council, 16149 Genoa, Italy; and 4 Department of Pharmacology, University of Minnesota at Minneapolis, Minneapolis, Minnesota 55455

Cyclic ADP-ribose (cADPR), a universal calcium releaser, is generated from NAD+ by an ADP-ribosyl cyclase and is degraded to ADP-ribose by a cADPR hydrolase. In mammals, both activities are expressed as ectoenzymes by the transmembrane glycoprotein CD38. CD38 was identified in both epithelial cells and smooth myocytes isolated from bovine trachea. Intact tracheal smooth myocytes (TSMs) responded to extracellular cADPR (100 µM) with an increase in intracellular calcium concentration ([Ca2+]i) both at baseline and after acetylcholine (ACh) stimulation. The nonhydrolyzable analog 3-deaza-cADPR (10 nM) elicited the same effects as cADPR, whereas the cADPR antagonist 8-NH2-cADPR (10 µM) inhibited both basal and ACh-stimulated [Ca2+]i levels. Extracellular cADPR or 3-deaza-cADPR caused a significant increase of ACh-induced contraction in tracheal smooth muscle strips, whereas 8-NH2-cADPR decreased it. Tracheal mucosa strips, by releasing NAD+, enhanced [Ca2+]i in isolated TSMs, and this increase was abrogated by either NAD+-ase or 8-NH2-cADPR. These data suggest the existence of a paracrine mechanism whereby mucosa-released extracellular NAD+ plays a hormonelike function and cADPR behaves as second messenger regulating calcium-related contractility in TSMs.

acetylcholine; CD38; nicotinamide adenine dinucleotide/ cyclic adenosine 5'-diphosphate-ribose-mediated paracrine mechanisms; calcium-related contraction of tracheal strips; adenosine 5'-diphosphate-ribosyl cyclase; cyclic adenosine 5'-diphosphate-ribose hydrolase.


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