Ventura S, Bavetta S, Milner P, Ralevic V, Burnstock G
Department of Anatomy and Developmental Biology, University College London, U.K.
Neuroscience. 1998 Mar;83(2):607-16. doi: 10.1016/s0306-4522(97)00416-8.
Cross-sections of the vas deferens taken from control adult male rats showed positive histochemical reactivity to acetylcholinesterase and immunoreactivity for antibodies to protein gene product 9.5, tyrosine hydroxylase, neuropeptide Y, vasoactive intestinal polypeptide, nitric oxide synthase and calcitonin gene-related peptide. Immunoreactivity to substance P was very sparse. Histochemical reactivity to acetylcholinesterase and immunoreactivity to vasoactive intestinal polypeptide and nitric oxide synthase was concentrated in the subepithelial lamina propria and inner smooth muscle layers. Complete surgical denervation resulting from transection of the nerve arising from the pelvic ganglion which supplies the vas deferens totally abolished the immunoreactivity to all of the antibodies tested as well as the histochemical reactivity to acetylcholinesterase. In sections of the prostatic end of the vas deferens taken from rats neonatally pretreated with capsaicin, immunoreactivity to calcitonin gene-related peptide and substance P was reduced by 75 and 83%, respectively. Immunoreactivity to neuropeptide Y, vasoactive intestinal polypeptide and nitric oxide synthase was similar in tissue sections taken from capsaicin-treated rats and those taken from control tissues. Pretreatment of rats with guanethidine or 6-hydroxydopamine decreased immunoreactivity to tyrosine hydroxylase and neuropeptide Y by 60-70%, but immunoreactivity to substance P, vasoactive intestinal polypeptide and nitric oxide synthase was unchanged, while immunoreactivity to calcitonin gene-related peptide and acetylcholinesterase staining was increased by guanethidine but not by 6-hydroxydopamine treatment. Triple labelling experiments showed nitric oxide synthase, vasoactive intestinal polypeptide and acetylcholinesterase all to be co-localized in some nerve fibres. These results indicate that the nitric oxide synthase contained in the nerve fibres innervating the rat vas deferens is unaffected by pretreatment of rats with capsaicin, 6-hydroxydopamine or guanethidine but is abolished by surgical denervation, of postganglionic parasympathetic, sympathetic and sensory nerves. Therefore it appears that nitric oxide synthase is co-localized with vasoactive intestinal polypeptide in the postganglionic parasympathetic nerves which innervate the rat vas deferens.
取自成年雄性对照大鼠的输精管横切片显示，其对乙酰胆碱酯酶具有阳性组织化学反应活性，对蛋白质基因产物9.5、酪氨酸羟化酶、神经肽Y、血管活性肠肽、一氧化氮合酶和降钙素基因相关肽的抗体具有免疫反应性。对P物质的免疫反应性非常稀少。对乙酰胆碱酯酶的组织化学反应活性以及对血管活性肠肽和一氧化氮合酶的免疫反应性集中在黏膜下层固有层和内层平滑肌层。由供应输精管的盆腔神经节发出的神经横断导致的完全手术去神经支配，完全消除了对所有测试抗体的免疫反应性以及对乙酰胆碱酯酶的组织化学反应活性。在取自新生期用辣椒素预处理的大鼠的输精管前列腺端切片中，对降钙素基因相关肽和P物质的免疫反应性分别降低了75%和83%。在取自辣椒素处理大鼠的组织切片和取自对照组织的切片中，对神经肽Y、血管活性肠肽和一氧化氮合酶的免疫反应性相似。用胍乙啶或6-羟基多巴胺预处理大鼠，使对酪氨酸羟化酶和神经肽Y的免疫反应性降低了60%-70%，但对P物质、血管活性肠肽和一氧化氮合酶的免疫反应性未改变，而胍乙啶处理使对降钙素基因相关肽的免疫反应性和乙酰胆碱酯酶染色增加，6-羟基多巴胺处理则未增加。三重标记实验表明，一氧化氮合酶、血管活性肠肽和乙酰胆碱酯酶都共定位于一些神经纤维中。这些结果表明，支配大鼠输精管的神经纤维中所含的一氧化氮合酶不受用辣椒素、6-羟基多巴胺或胍乙啶预处理大鼠的影响，但节后副交感神经、交感神经和感觉神经的手术去神经支配可将其消除。因此，似乎一氧化氮合酶与血管活性肠肽在支配大鼠输精管的节后副交感神经中共定位。
