Smyth L M, Breen L T, Yamboliev I A, Mutafova-Yambolieva V N
Department of Physiology and Cell Biology, Anderson Medical Sciences Building/MS 352, University of Nevada School of Medicine, Reno, NV 89557-0271, USA.
Neuroscience. 2006;139(4):1467-77. doi: 10.1016/j.neuroscience.2006.01.043. Epub 2006 Mar 31.
Using high performance liquid chromatography fraction analysis we have recently established that numerous smooth muscle preparations, including the canine mesenteric artery and vein, release beta-nicotinamide adenine dinucleotide upon short-pulse electrical field stimulation in tetrodotoxin- and omega-conotoxin GVIA-sensitive manners [ Release of beta-nicotinamide adenine dinucleotide upon stimulation of postganglionic nerve terminals in blood vessels and urinary bladder. J Biol Chem 279:48893-48903.]. The beta-nicotinamide adenine dinucleotide metabolites ADP-ribose and cyclic ADP-ribose are also present in the tissue superfusates. CD38 is a multifunctional enzyme involved in the degradation of beta-nicotinamide adenine dinucleotide to ADP-ribose and cyclic ADP-ribose. Western immunoblot analysis revealed that CD38 is expressed in both artery and vein. Confocal laser scanning microscopy established colocalization of CD38 with tyrosine hydroxylase, synaptotagmin and synaptic vesicle protein in both blood vessels. High performance liquid chromatography with fluorescence detection demonstrated that whole tissue segments metabolize 1,N(6)-etheno-nicotinamide adenine dinucleotide to 1,N(6)-etheno-ADP-ribose and nicotinamide-guanine dinucleotide to cyclic GDP-ribose, suggesting the presence of both nicotinamide adenine dinucleotide-glycohydrolase and ADP-ribosyl cyclase activities in these blood vessels. Both enzymes appear to be associated with the membrane fraction, and therefore might be attributed to CD38. These data demonstrate a previously uncharacterized localization of CD38 in perivascular autonomic nerve terminals. Therefore, the beta-nicotinamide adenine dinucleotide/CD38 system may provide new mechanisms in autonomic neurovascular control.
利用高效液相色谱分级分析，我们最近证实，包括犬肠系膜动脉和静脉在内的多种平滑肌制剂，在河豚毒素和ω-芋螺毒素GVIA敏感的方式下，经短脉冲电场刺激后会释放β-烟酰胺腺嘌呤二核苷酸[血管和膀胱节后神经末梢受刺激后β-烟酰胺腺嘌呤二核苷酸的释放。《生物化学杂志》279:48893 - 48903。]。组织灌流液中也存在β-烟酰胺腺嘌呤二核苷酸代谢物ADP-核糖和环ADP-核糖。CD38是一种多功能酶，参与将β-烟酰胺腺嘌呤二核苷酸降解为ADP-核糖和环ADP-核糖。蛋白质免疫印迹分析显示，CD38在动脉和静脉中均有表达。共聚焦激光扫描显微镜证实，在两种血管中，CD38与酪氨酸羟化酶、突触结合蛋白和突触小泡蛋白共定位。高效液相色谱荧光检测表明，整个组织片段可将1,N(6)-乙烯基烟酰胺腺嘌呤二核苷酸代谢为1,N(6)-乙烯基-ADP-核糖，将烟酰胺-鸟嘌呤二核苷酸代谢为环GDP-核糖，这表明这些血管中存在烟酰胺腺嘌呤二核苷酸糖水解酶和ADP-核糖基环化酶活性。这两种酶似乎都与膜部分相关，因此可能归因于CD38。这些数据证明了CD38在血管周围自主神经末梢中以前未被描述的定位。因此，β-烟酰胺腺嘌呤二核苷酸/CD38系统可能为自主神经血管控制提供新机制。
