Yu J G, Kimura T, Chang X F, Lee T J
Department of Pharmacology, Southern Illinois University School of Medicine, P.O. Box 19230, Springfield, IL 62794-1222, USA.
Brain Res. 1998 Aug 10;801(1-2):78-87. doi: 10.1016/s0006-8993(98)00548-4.
The distribution of nitric oxide synthase (NOS)-, choline acetyltransferase (ChAT)-, and vasoactive intestinal polypeptide (VIP)-immunoreactivities, and nicotinamide adenine dinucleotide phosphate diaphorase (NADPHd)-reactivities in the sphenopalatine ganglia (SPG), and perivascular nerves in middle cerebral arteries of the pig was investigated by double-staining techniques using combined immunofluorescence and histochemistry methods. In the SPG, almost all ganglionic cells were NOS-immunoreactive (I) and NADPHd-positive, and both NOS immunoreactivities and NADPHd reactivities were completely co-localized. ChAT-I ganglionic cells accounted for 75%, while VIP-I ganglionic cells represented 42% of all ganglionic cells. Almost all VIP immunoreactivities were co-localized with ChAT immunoreactivities, and all ganglionic cells that were VIP-I and/or ChAT-I were NOS-I and NADPHd-reactive. None of the ganglionic cells in the SPG were immunoreactive to calcitonin gene-related peptide (CGRP). CGRP immunoreactivities, however, were found to surround some ganglionic cells. In middle cerebral arteries, all adventitial NOS-I bundles and fine fibers were coincident with NADPHd fibers. Almost all adventitial ChAT-I bundles and thin fibers, and VIP-I mesh-like fibers stained positively for NADPHd, while the mesh-like NADPHd fine fibers were not ChAT-I. Simultaneous labeling using antibodies against VIP and ChAT further indicated that VIP-I fibers were closer than ChAT-I fibers to the smooth muscle. In rare occasions, perivascular fibers were found to be stained for both ChAT and VIP, showing that most ChAT-I and VIP-I fibers were not coincident. These results suggest that ChAT and VIP are rarely co-localized in perivascular nerves in middle cerebral arteries, and point out that the neurotransmitter and the modulator that are co-localized within the same nerve cell body may distribute totally independently and differently at the terminal level. The present results also indicate that in cerebral perivascular nerves, the combination of nitric oxide (NO) and acetylcholine (ACh), as well as the combination of NO and VIP, are localized in the same nerve with different axons containing either NO plus ACh, or NO plus VIP. These findings support the hypothesis that ACh and VIP may act as modulators in regulating presynaptic release of NO, and therefore, cerebral neurogenic vasodilation, from their respective perivascular cholinergic-nitric oxidergic and VIPergic-nitric oxidergic nerves.
采用免疫荧光和组织化学相结合的双重染色技术，研究了猪蝶腭神经节（SPG）以及大脑中动脉血管周围神经中一氧化氮合酶（NOS）、胆碱乙酰转移酶（ChAT）和血管活性肠肽（VIP）免疫反应性及烟酰胺腺嘌呤二核苷酸磷酸黄递酶（NADPHd）反应性的分布情况。在SPG中，几乎所有神经节细胞均为NOS免疫反应性（I）且NADPHd阳性，NOS免疫反应性和NADPHd反应性完全共定位。ChAT-I神经节细胞占所有神经节细胞的75%，而VIP-I神经节细胞占42%。几乎所有VIP免疫反应性均与ChAT免疫反应性共定位，所有VIP-I和/或ChAT-I神经节细胞均为NOS-I且具有NADPHd反应性。SPG中没有神经节细胞对降钙素基因相关肽（CGRP）呈免疫反应性。然而，发现CGRP免疫反应性围绕着一些神经节细胞。在大脑中动脉中，所有外膜NOS-I束和细纤维与NADPHd纤维一致。几乎所有外膜ChAT-I束和细纤维以及VIP-I网状纤维NADPHd染色呈阳性，而网状NADPHd细纤维不是ChAT-I。使用抗VIP和ChAT抗体的同时标记进一步表明，VIP-I纤维比ChAT-I纤维更靠近平滑肌。在极少数情况下，发现血管周围纤维ChAT和VIP均染色，表明大多数ChAT-I和VIP-I纤维不重合。这些结果表明，ChAT和VIP在大脑中动脉血管周围神经中很少共定位，并指出在同一神经细胞体内共定位的神经递质和调节剂在终末水平可能完全独立且不同地分布。目前的结果还表明，在脑血管周围神经中，一氧化氮（NO）和乙酰胆碱（ACh）的组合以及NO和VIP的组合，位于同一神经中，不同轴突分别含有NO加ACh或NO加VIP。这些发现支持以下假设：ACh和VIP可能作为调节剂调节各自血管周围胆碱能-一氧化氮能神经和VIP能-一氧化氮能神经的突触前NO释放，从而调节脑血管源性舒张。
