Differential expression and induction of mRNAs encoding two inducible nitric oxide synthases in rat kidney.
We used quantitative PCR methods and renal microdissection to characterize the expression of inducible nitric oxide synthase (iNOS) mRNAs in rat kidney and cultured glomerular mesangial cells. A partial cDNA homologous to murine macrophage iNOS (macNOS), but distinct from rat vascular smooth muscle iNOS (vsmNOS), was cloned from normal rat kidney. macNOS was the principal iNOS isoform tonically expressed in microdissected glomeruli, proximal tubules, medullary thick ascending limbs (mTAL), cortical and inner medullary collecting ducts (IMCD), and cultured mesangial cells, whereas vsmNOS was the major isoform expressed in arcuate and interlobular arteries. Basal macNOS expression was greatest in mTALs and IMCDs. Restriction mapping of RT-PCR products indicated that basal expression of macNOS mRNA was comparable to that of vsmNOS in cortex, but greater than vsmNOS in outer and inner medulla. However, compared to controls, lipopolysaccharide (LPS)-treated rats exhibited a much greater proportion of vsmNOS mRNA and higher levels of total iNOS mRNA in each zone. Similarly, TNF alpha and IF-gamma preferentially induced expression of vsmNOS mRNA in cultured mesangial cells. We conclude that two iNOS isoforms are constitutively and heterogeneously expressed in the normal rat kidney, and that endotoxemia and cytokines differentially induce their expression.
Mohaupt
M G
MG
DCI Laboratory of Molecular Biology in Nephrology, University of Florida College of Medicine, Gainesville.
Elzie
J L
JL
Ahn
K Y
KY
Clapp
W L
WL
Wilcox
C S
CS
Kone
B C
BC
D000594:Amino Acid Oxidoreductases / Q000096:biosynthesis* / Q000235:genetics*; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002478:Cells, Cultured; D016207:Cytokines / Q000494:pharmacology; D017931:DNA Primers; D004790:Enzyme Induction / Q000187:drug effects; D015971:Gene Expression Regulation, Enzymologic; D005920:Glomerular Mesangium / Q000201:enzymology*; D007527:Isoenzymes / Q000096:biosynthesis / Q000235:genetics; D007684:Kidney Tubules / Q000201:enzymology*; D008070:Lipopolysaccharides / Q000494:pharmacology; D008297:Male; D008969:Molecular Sequence Data; D019001:Nitric Oxide Synthase; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000096:biosynthesis*; D051381:Rats; D017207:Rats, Sprague-Dawley
D016207:Cytokines; D017931:DNA Primers; D007527:Isoenzymes; D008070:Lipopolysaccharides; D012333:RNA, Messenger; D019001:Nitric Oxide Synthase; D000594:Amino Acid Oxidoreductases