INDUCTION AND CDNA SEQUENCE OF INDUCIBLE NITRIC-OXIDE SYNTHASE FROM CANINE AORTIC SMOOTH-MUSCLE CELLS

Citation
Xf. Wang et al., INDUCTION AND CDNA SEQUENCE OF INDUCIBLE NITRIC-OXIDE SYNTHASE FROM CANINE AORTIC SMOOTH-MUSCLE CELLS, American journal of physiology. Heart and circulatory physiology, 44(4), 1998, pp. 1122-1129
Citations number
29
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Physiology
ISSN journal
0363-6135
Volume
44
Issue
4
Year of publication
1998
Pages
1122 - 1129
Database
ISI
SICI code
0363-6135(1998)44:4<1122:IACSOI>2.0.ZU;2-#
Abstract
An inducible isoform of nitric oxide synthase (type II, iNOS) is expre ssed in cardiac and vascular smooth muscle in response to inflammatory cytokines. The dog is an important large animal used for cardiovascul ar research including effects of exercise, heart failure, and allograf t rejection. However, molecular probes for iNOS developed in other mam mals have not been reliable for the study of iNOS induction in canine vascular smooth muscle. Experiments were designed to develop a molecul ar probe for canine iNOS. Smooth muscle cells were isolated from canin e aortas. The cells (passages 3-10) were incubated for 1, 3, 6, 12, 24 , 48, or 72 h in the absence and presence of Escherichia coli lipopoly saccharide (LPS) to induce iNOS. Total RNA was isolated from the cells using standard techniques. RT-PCR with primers against conserved regi ons of all known iNOS enzyme was used to clone the iNOS cDNA. RT-PCR s howed a single band only from cells treated with LPS. Cloned cDNA from cultured canine aortic smooth muscle cells has 84% homology to human, 81% to rat, and 81% to mouse iNOS gene. Identification of the cDNA fo r canine iNOS will be useful in the study of differential, transcripti onal regulation of inducible (type II) compared with constitutive endo thelial (type III) NOS in canine studies of allograft rejection and ca rdiovascular disease.