Ultrastructural localization of nitric oxide synthase in canine small intestine and colon.
The ultrastructural distribution and subcellular localization of nitric oxide synthase (NOS) immunoreactivity and its possible colocalization with vasoactive intestinal polypeptide (VIP) and substance P in the muscularis externa in canine ileum and colon were studied by using polyclonal antisera raised against VIP, substance P, and cerebellar NOS. Immunogold staining, with or without silver enhancement, was carried out directly on ultrathin sections using single and two-faced double immunogold methods. NOS immunoreactivity was observed in nerve profiles in myenteric plexus and circular muscle layer. Immunoreactivity was occasionally detected in smooth muscle cells and interstitial cells of Cajal. The double immunostaining revealed NOS and VIP in the same nerve varicosities but never in the same organelles. NOS was localized in electron-dense material of undetermined nature, whereas VIP was associated with large granular vesicles. Substance P and NOS were never found in the same nerves. These results indicate that NOS is present in the enteric nerves containing VIP but in different organelles and that nitric oxide release probably does not occur by an exocytotic mechanism.
Berezin
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Department of Biomedical Sciences, McMaster University, Hamilton, Ontario, Canada.
Snyder
S H
SH
Bredt
D S
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Daniel
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EE
D000594:Amino Acid Oxidoreductases / Q000378:metabolism*; D000818:Animals; D003106:Colon / Q000294:innervation / Q000378:metabolism* / Q000648:ultrastructure; D004285:Dogs; D007150:Immunohistochemistry; D007421:Intestine, Small / Q000294:innervation / Q000378:metabolism* / Q000648:ultrastructure; D016253:Microscopy, Immunoelectron; D009420:Nervous System / Q000378:metabolism; D019001:Nitric Oxide Synthase; D013194:Staining and Labeling; D013373:Substance P / Q000378:metabolism; D014018:Tissue Distribution; D014660:Vasoactive Intestinal Peptide / Q000378:metabolism
D013373:Substance P; D014660:Vasoactive Intestinal Peptide; D019001:Nitric Oxide Synthase; D000594:Amino Acid Oxidoreductases