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Immunocytochemical localization of distinct isoforms of nitric oxide synthase in the juxtaglomerular apparatus of normal rat kidney.

A Tojo, S S Gross, L Zhang, C C Tisher, H H Schmidt, C S Wilcox and K M Madsen
JASN January 1994, 4 (7) 1438-1447;
A Tojo
Division of Nephrology, University of Florida, Gainesville 32610-0224.
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S S Gross
Division of Nephrology, University of Florida, Gainesville 32610-0224.
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L Zhang
Division of Nephrology, University of Florida, Gainesville 32610-0224.
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C C Tisher
Division of Nephrology, University of Florida, Gainesville 32610-0224.
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H H Schmidt
Division of Nephrology, University of Florida, Gainesville 32610-0224.
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C S Wilcox
Division of Nephrology, University of Florida, Gainesville 32610-0224.
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K M Madsen
Division of Nephrology, University of Florida, Gainesville 32610-0224.
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Abstract

Nitric oxide (NO) is a messenger molecule that functions as a vasodilator and accounts for the biologic activity of endothelium-derived relaxing factor (EDRF). The enzyme responsible for NO production, NO synthase (NOS), exists in different isoforms. Some are expressed constitutively in various tissues, whereas others require induction by endotoxin and cytokines. There is evidence that NO plays an important role in the regulation of basal renal vascular resistance and in the tubuloglomerular feedback response. Specific antibodies to a constitutive NOS isolated from rat brain (B-NOS) and an inducible NOS isolated from rat aortic smooth muscle (VSM-NOS) were used to establish the cellular and subcellular distribution of NOS in the kidney. Kidneys from normal rats were preserved and processed for light and electron microscopic immunohistochemical studies using both a preembedding and a postembedding horseradish peroxidase technique. By light microscopy, strong immunostaining for B-NOS was observed in the juxtaglomerular apparatus, where it was located in cells of the macula densa. In contrast, immunostaining for VSM-NOS was specifically located in the terminal afferent arteriole and occasionally also in the initial efferent arteriole in the juxtaglomerular apparatus. In addition, faint immunostaining was present in the entire distal tubule. There was no labeling of arcuate or interlobular arteries. The injection of lipopolysaccharide was associated with increased immunostaining for VSM-NOS in the afferent arteriole but had no effect on B-NOS staining. By electron microscopy, B-NOS immunostaining was present throughout the cytoplasm of the macula densa cells, where it appeared to be associated mainly with small vesicles. Immunostaining for VSM-NOS in the afferent arteriole exhibited a patchy distribution in some cells, whereas other cells were stained throughout the cytoplasm. These results indicate that two distinct isoforms of NOS are present in the juxtaglomerular apparatus and support the hypothesis that NOS is involved in the regulation of tubuloglomerular feedback and glomerular capillary pressure.

  • Copyright © 1994 by American Society of Nephrology
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Journal of the American Society of Nephrology
Vol. 4, Issue 7
1 Jan 1994
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Immunocytochemical localization of distinct isoforms of nitric oxide synthase in the juxtaglomerular apparatus of normal rat kidney.
A Tojo, S S Gross, L Zhang, C C Tisher, H H Schmidt, C S Wilcox, K M Madsen
JASN Jan 1994, 4 (7) 1438-1447;

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Immunocytochemical localization of distinct isoforms of nitric oxide synthase in the juxtaglomerular apparatus of normal rat kidney.
A Tojo, S S Gross, L Zhang, C C Tisher, H H Schmidt, C S Wilcox, K M Madsen
JASN Jan 1994, 4 (7) 1438-1447;
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  • Effect of Graded Hypoxia on the Induction and Function of Inducible Nitric Oxide Synthase in Rat Mesangial Cells
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