In situ hybridization localization of mRNA encoding inducible nitric oxide synthase in rat kidney.
We used in situ hybridization with a digoxigenin-labeled cRNA for inducible nitric oxide synthase (iNOS) to characterize the intrarenal distribution of iNOS transcripts in normal and lipopolysaccharide (LPS)-treated rats. In normal rats, the S3 segment of the proximal tubule, the cortical and medullary thick ascending limb, the distal convoluted tubule, and the cortical and inner medullary collecting duct were intensely labeled, whereas the thin limbs of Henle, proximal convoluted tubule, outer medullary collecting duct, and medullary interstitial cells were weakly labeled. LPS-treated rats exhibited a similar labeling pattern, but with increased staining of mesangial cells, medullary interstitial cells, and papillary surface epithelium. The renal vasculature, including the afferent arteriole, was not labeled in either group. No cellular labeling was observed when the sections were hybridized with the sense iNOS probe. These results indicate that iNOS mRNA is tonically and differentially expressed along the normal rat nephron and that LPS induces iNOS gene expression in normally quiescent mesangial cells, medullary interstitial cells, and papillary surface epithelium.
Ahn
K Y
KY
DCI Laboratory of Molecular Biology in Nephrology, University of Florida College of Medicine, Gainesville 32610-0224.
Mohaupt
M G
MG
Madsen
K M
KM
Kone
B C
BC
D000594:Amino Acid Oxidoreductases / Q000096:biosynthesis*; D000818:Animals; D015152:Blotting, Northern; D002460:Cell Line; D004790:Enzyme Induction; D004848:Epithelium / Q000187:drug effects / Q000201:enzymology / Q000648:ultrastructure; D015870:Gene Expression; D017403:In Situ Hybridization / Q000379:methods; D007668:Kidney / Q000187:drug effects / Q000201:enzymology* / Q000648:ultrastructure; D007672:Kidney Cortex / Q000201:enzymology; D007679:Kidney Medulla / Q000201:enzymology; D007686:Kidney Tubules, Distal / Q000201:enzymology; D007687:Kidney Tubules, Proximal / Q000201:enzymology; D008070:Lipopolysaccharides / Q000494:pharmacology; D008264:Macrophages; D008297:Male; D051379:Mice; D019001:Nitric Oxide Synthase; D015347:RNA Probes; D018075:RNA, Complementary / Q000032:analysis; D012333:RNA, Messenger / Q000032:analysis* / Q000096:biosynthesis; D051381:Rats; D017207:Rats, Sprague-Dawley; D012016:Reference Values
D008070:Lipopolysaccharides; D015347:RNA Probes; D018075:RNA, Complementary; D012333:RNA, Messenger; D019001:Nitric Oxide Synthase; D000594:Amino Acid Oxidoreductases