Polymerase chain reaction analysis of renin in rat aortic smooth muscle.
Controversy exists whether vascular smooth muscle cells in vivo synthesize renin, thereby providing a critical component of the hypothesized vascular renin-angiotensin system. To examine this question, we enzymatically isolated and pooled the medial layer of thoracic aortas from Sprague-Dawley rats that were either untreated or enalapril treated for 3 days, isolated messenger RNA (mRNA), and performed Northern blot analysis with rat complementary DNA (cDNA) probes for renin, cathepsin D, and cathepsin E. Renin mRNA was detected in kidney but was not detected in aortic smooth muscle from the untreated or enalapril-treated groups. Cathepsin E mRNA was detected in enalapril-treated aorta and kidney, and cathepsin D mRNA was detected in all tissues examined. cDNA was synthesized and subjected to polymerase chain reaction analysis by using primers corresponding in sequence to regions conserved throughout the aspartic proteinases. Cathepsins D and E were amplified from kidney and aortic cDNA. Renin was less consistently amplified from the aortic cDNA and was much less abundant than cathepsin E or cathepsin D. These results suggest that 1) renin mRNA is present in aortic smooth muscle cells in vivo in quantities detectable only after multiple rounds of polymerase chain reaction amplification, 2) renin mRNA is not upregulated in aortic smooth muscle after converting enzyme inhibition, and 3) cathepsins D and E are the predominant aspartic proteinases in aortic smooth muscle.
Holycross
B J
BJ
Department of Pharmacology, University of Virginia School of Medicine, Charlottesville.
Saye
J
J
Harrison
J K
JK
Peach
M J
MJ
D000818:Animals; D001011:Aorta / Q000378:metabolism*; D001483:Base Sequence; D002403:Cathepsins / Q000235:genetics; D004247:DNA / Q000096:biosynthesis; D004656:Enalapril / Q000494:pharmacology; D005954:Glucosephosphate Dehydrogenase / Q000235:genetics; D008297:Male; D008969:Molecular Sequence Data; D009131:Muscle, Smooth, Vascular / Q000378:metabolism*; D015345:Oligonucleotide Probes / Q000235:genetics; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D011919:Rats, Inbred Strains; D012083:Renin / Q000235:genetics / Q000378:metabolism*; D014335:Tropomyosin / Q000235:genetics
D015345:Oligonucleotide Probes; D012333:RNA, Messenger; D014335:Tropomyosin; D004656:Enalapril; D004247:DNA; D005954:Glucosephosphate Dehydrogenase; D002403:Cathepsins; D012083:Renin