title: Release of beta-nicotinamide adenine dinucleotide upon stimulation of postganglionic nerve terminals in blood vessels and urinary bladder.
abstract: Chemical signaling in autonomic neuromuscular transmission involves agents that function as neurotransmitters and/or neuromodulators. Using high performance liquid chromatography techniques with fluorescence and electrochemical detection we observed that, in addition to ATP and norepinephrine (NE), electrical field stimulation (EFS, 4-16 Hz, 0.1-0.3 ms, 15 V, 60-120 s) of isolated vascular and non-vascular preparations co-releases a previously unidentified compound with apparent nucleotide or nucleoside structure. Extensive screening of more than 25 nucleotides and nucleosides followed by detailed peak identification revealed that beta-nicotinamide adenine dinucleotide (beta-NAD) is released in tissue superfusates upon EFS of canine mesenteric artery (CMA), canine urinary bladder, and murine urinary bladder in the amounts of 7.1 +/- 0.7, 26.5 +/- 4.5, and 15.1 +/- 3.2 fmol/mg of tissue, respectively. Smaller amounts of the beta-NAD metabolites cyclic adenosine 5'-diphosphoribose (cADPR) and ADPR were also present in the superfusates collected during EFS of CMA (2.5 +/- 0.9 and 5.8 +/- 0.8 fmol/mg of tissue, respectively), canine urinary bladder (1.8 +/- 0.5 and 9.0 +/- 6.0 fmol/mg of tissue, respectively), and murine urinary bladder (1.4 +/- 0.1 and 6.2 +/- 2.4 fmol/mg of tissue, respectively). The three nucleotides were also detected in the samples collected before EFS (0.2-1.6 fmol/mg of tissue). Exogenous beta-NAD, cADPR, and ADPR (all 100 nm) reduced the release of NE in CMA at 16 Hz from 27.8 +/- 6.0 fmol/mg of tissue to 15.5 +/- 5.0, 12 +/- 3.0, and 10.0 +/- 4.0 fmol/mg of tissue, respectively. In conclusion, we detected constitutive and nerve-evoked overflow of beta-NAD, cADPR, and ADPR in vascular and non-vascular smooth muscles, beta-NAD being the prevailing compound. These substances modulate the release of NE, implicating novel nucleotide mechanisms of autonomic nervous system control of smooth muscle.
authors.affiliation: Department of Physiology and Cell Biology, University of Nevada School of Medicine, Reno, Nevada 89557-0271, USA.
authors.forename: Lisa M
authors.initials: LM
authors.lastname: Smyth
authors.forename: Janette
authors.initials: J
authors.lastname: Bobalova
authors.forename: Michael G
authors.initials: MG
authors.lastname: Mendoza
authors.forename: Christy
authors.initials: C
authors.lastname: Lew
authors.forename: Violeta N
authors.initials: VN
authors.lastname: Mutafova-Yambolieva
mesh_terms: D000079:Acetaldehyde / Q000031:analogs & derivatives* / Q000737:chemistry; D000225:Adenine / Q000737:chemistry; D000255:Adenosine Triphosphate / Q000737:chemistry; D018663:Adrenergic Agents / Q000494:pharmacology; D000779:Anesthetics, Local / Q000494:pharmacology; D000818:Animals; D002121:Calcium Channel Blockers / Q000494:pharmacology; D002851:Chromatography, High Pressure Liquid; D036563:Cyclic ADP-Ribose / Q000737:chemistry; D004285:Dogs; D004563:Electrochemistry; D004594:Electrophysiology; D005260:Female; D006145:Guanethidine / Q000494:pharmacology; D006168:Guinea Pigs; D006863:Hydrogen-Ion Concentration; D008297:Male; D008638:Mesenteric Arteries / Q000378:metabolism*; D051379:Mice; D008956:Models, Chemical; D009130:Muscle, Smooth / Q000378:metabolism; D009243:NAD / Q000378:metabolism*; D009638:Norepinephrine / Q000737:chemistry; D016627:Oxidopamine / Q000494:pharmacology; D011817:Rabbits; D051381:Rats; D017208:Rats, Wistar; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013314:Stress, Mechanical; D013696:Temperature; D013779:Tetrodotoxin / Q000494:pharmacology; D013997:Time Factors; D001743:Urinary Bladder / Q000378:metabolism*; D020866:omega-Conotoxin GVIA / Q000494:pharmacology
chemical_list: D018663:Adrenergic Agents; D000779:Anesthetics, Local; D002121:Calcium Channel Blockers; D009243:NAD; D036563:Cyclic ADP-Ribose; D013779:Tetrodotoxin; D016627:Oxidopamine; D000255:Adenosine Triphosphate; D020866:omega-Conotoxin GVIA; C004656:chloroacetaldehyde; D000079:Acetaldehyde; D000225:Adenine; D009638:Norepinephrine; D006145:Guanethidine