Abstract
The localization of nitric oxide synthase (NOS) in vascular endothelial cells of submucosal blood vessels from the guinea-pig ileum was examined using NADPH diaphorase histochemistry at the light microscopic level, and endothelial NOS immunohistochemistry at the light and electron microscopic level. The pattern of staining observed following NADPH diaphorase histochemistry and endothelial NOS immunohistochemistry was identical. Endothelial cells of the arterioles, capillaries and venules showed small patches of intense, perinuclear staining. Under the electron microscope, endothelial NOS immunoreactivity was found predominantly in association with the Golgi apparatus and with the membranes of some vesicles. Small regions of the plasma membrane and the rough endoplasmic reticulum also showed some immunoreactivity. The presence of NOS in the Golgi apparatus and in vesicles raises the possibility that NOS may be exteriorized by endothelial cells, and hence that nitric oxide is synthesized extracellularly.
Similar content being viewed by others
References
Alm P, Larsson B, Ekblad E, Sundler F, Andersson K-E (1993) Immunohistochemical localization of peripheral nitric oxide synthase-containing nerves using antibodies raised against synthesized C-and N-terminal fragments of a cloned enzyme from rat brain. Acta Physiol Scand 148:421–429
Anderson CR, Furness JB, Woodman HL, Edwards SL, Crack P, Smith AI (1995) Characterization of enteric neurons with nitric oxide synthase immunoreactivity that project to prevertebral ganglia. J Auton Nerv Syst (in press)
Bandaletova T, Brouet I, Bartsch H, Sugimura T, Esumi H, Ohshima H (1993) Immunohistochemical localization of an inducible form of nitric oxid synthase in various organs of rats treated withPropionibacterium acnes and lipopolysaccarides. Acta Pathol Microbiol Immunol Scand (APMIS) 101:330–336
Boje KM, Fung H (1990) Endothelial nitric oxide generating enzyme(s) in the bovine aorta: subcellular location and metabolic characterization. J Pharmacol Exp Ther 253: 20–26
Bredt DS, Snyder SH (1992) Nitric oxide, a novel neuronal messenger. Neuron 8:3–11
Bredt DS, Hwang PM, Snyder SH (1990) Localization of nitric oxide synthase indicating a neural role for nitric oxide. Nature 347:768–770
Busconi L, Michel T (1993) Endothelial nitric oxide synthase. N-terminal myristoylation determines subcellular localization. J Biol Chem 268:8410–8413
Eldred WD, Zucker C, Karten HJ, Yazulla S (1983) Comparison of fixation and penetration enhancement techniques for use in ultrastructural immunocytochemistry. J Histochem Cytochem 31:285–292
Förstermann U, Schmidt HHHW, Pollock JS, Sheng H, Mitchell JA, Warner TD, Nakane M, Murad F (1991) Isoforms of nitric oxide synthase. Biochem Pharmacol 42:1849–1859
Furchgott RF, Zawadzki JV (1980) The obligatory role of endothelial cells in the relaxation of arterial smooth muscle by acetylcholine. Nature 288:373–376
Hecker M, Mülsch A, Busse R (1994a) Subcellular localization and characterization of neuronal nitric oxide synthase. J Neurochem 62:1524–1529
Hecker M, Mülsch A, Bassenge E, Förstermann U, Busse R (1994b) Subcellular localization and characterization of nitric oxide synthase(s) in endothelial cells: physiological implications. Biochem J 299:247–252
Hibbs JB, Taintor RR, Vavrin Z, Rachlin EM (1988) Nitric oxide: a cytotoxic activated macrophage effector molecule. Biochem Biophys Res Commun 157:87–94
Ignarro LJ (1989) Biological actions and properties of endothelium-derived nitric oxide formed and released from artery and vein. Circ Res 65:1–21
Lamas S, Marsden PA, Li GK, Tempst P, Michel T (1992) Endothelial nitric oxid synthase: molecular cloning and characterization of a distinct constitutive enzyme isoform. Proc Natl Acad Sci USA 89:6348–6352
Llewellyn-Smith IJ, Costa M, Furness JB (1985) Light and electron microscopic immunocytochemistry of the same nerves from whole mount preparations. J Histochem Cytochem 33:857–866
Loesch A, Belai A, Burnstock G (1994) An ultrastructural study of NADPH-diaphorase and nitric oxide synthase in the perivascular nerves and vascular endothelium of the rat basilar artery. J Neurocytol 23:49–59
Marletta MA (1993) Nitric oxide synthase structure and mechanism. J Biol Chem 268:12231–12234
Nichols K, Krantis A, Staines W (1992) Histochemical localization of nitric oxide-synthesizing neurons and vascular sites in the guinea-pig intestine. Neuroscience 51:791–799
Nichols K, Staines W, Krantis A (1993) Nitric oxide synthase distribution in the rat intestine: a histochemical analysis. Gastroenterology 105:1651–1661
Nichols K, Staines W, Rubin S, Krantis A (1994) Distribution of nitric oxide synthase activity in arterioles and venules of rat and human intestine. Am J Physiol 267:G270-G275
Nishida K, Harrison DG, Navas JP, Fisher AA, Dockery SP, Uematsu M, Nerem RM, Alexander RW, Murphy TJ (1992) Molecular cloning and characterization of the constitutive bovine aortic endothelial cell nitric oxide synthase. J Clin Invest 90:2092–2096
Palmer RMJ, Ferrige AG, Moncada S (1987) Nitric oxide release accounts for the biological activity of endothelium-derived relaxing factor. Nature 327:524–526
Poeggel G, Muller M, Seidel I, Rechardt L, Bernstein H-G (1993) Histochemistry of guanylate cyclase, phosphodiesterase and NADPH diaphorase (nitric oxide synthase) in rat brain vasculature. J Cardiovasc Pharmacol 20 [Suppl 12]:S76-S79
Pollock JS, Klinghofer V, Förstermann U, Murad F (1992) Endothelial nitric oxide synthase is myristylated. FEBS Lett 309:402–404
Pollock JS, Nakane M, Buttery LDK, Martinez A, Springall DR, Polak JM, Förstermann U, Murad F (1993) Characterization and localization of endothelial nitric oxide synthase using specific monoclonal antibodies. Am J Physiol 265:C1379-C1387
Scherer-Singler U, Vincent SR, Kimura H, McGeer EG (1983) Demonstration of a unique population of neurons with NADPH-diaphorase histochemistry. J Neurosci Methods 9:229–234
Sessa WC, Barber CM, Lynch KR (1993) Mutation of N-myristoylation site converts endothelial cell nitric oxide synthase from a membrane to a cytosolic protein. Circ Res 72:921–924
Spicer SS, Spivey MA, Ito M, Schulte BA (1994) Some ascites monoclonal antibody preparations contain contaminants that bind to selected Golgi zones or mast cells. J Histochem Cytochem 42:213–221
Towler DA, Eubanks SR, Towery DS, Adams SP, Glaser L (1987) Amino-terminal processing of proteins by N-myristoylation. Substrate specificity of N-myristoyl transferase. J Biol Chem 262:1030–1036
Tracey WR, Nakane M, Pollock JS, Förstermann U (1993) Nitric oxide synthases in neuronal cells, macrophages and endothelium are NADPH diaphorases, but represent only a fraction of total cellular NADPH diaphorase activity. Biochem Biophys Res Commun 195:1035–1040
Valtschanoff JG, Weinberg RJ, Rustioni A (1992) NADPH diaphorase in the spinal cord of rats J Comp Neurol 321:209–222
Vincent SR, Kimura H (1992) Histochemical mapping of nitric oxide synthase in the rat brain. Neuroscience 46:755–784
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
O'Brien, A.J., Young, H.M., Povey, J.M. et al. Nitric oxide synthase is localized predominantly in the Golgi apparatus and cytoplasmic vesicles of vascular endothelial cells. Histochem Cell Biol 103, 221–225 (1995). https://doi.org/10.1007/BF01454027
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF01454027