Cloning and expression of rat prostaglandin endoperoxide synthase (cyclooxygenase)-2 cDNA.
Rat prostaglandin endoperoxidase synthase-2 (PGHS-2) cDNA was cloned from rat calvarial osteoblasts total RNA by RT-PCR. The primary sequence of rat PGHS-2 had 98% and 92% identity to the mouse and human enzymes, respectively. Transfection of the rat PGHS-2 cDNA into COS 7 cells, followed by the addition of 20 microM arachidonic acid, resulted in a dramatic increase in PGE2 released from these cells. The amount of PGE2 produced was comparable to that obtained from cells similarly transfected with human PGHS-1 cDNA. In the rat paw carrageenin-oedema inflammatory model, the injected paw had elevated levels of PGHS-2 mRNA compared to the control paw. In a rat pyrexia model, injection of the pyrogen lipopolysaccharide, resulted in elevated levels of PGHS-2 mRNA in the brain. These results suggest that PGHS-2 expression plays a role both in inflammation and fever.
Kennedy
B P
BP
Department of Molecular Biology, Merck Frosst Centre for Therapeutic Research, Merck Frosst Canada Inc., Pointe Claire-Dorval, Quebec.
Chan
C C
CC
Culp
S A
SA
Cromlish
W A
WA
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D002351:Carrageenan; D002460:Cell Line; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000378:metabolism*; D015232:Dinoprostone / Q000378:metabolism; D004487:Edema / Q000139:chemically induced / Q000201:enzymology; D004790:Enzyme Induction; D015870:Gene Expression; D006801:Humans; D007249:Inflammation / Q000201:enzymology; D008070:Lipopolysaccharides / Q000494:pharmacology; D008297:Male; D051379:Mice; D008969:Molecular Sequence Data; D010006:Osteoblasts / Q000378:metabolism*; D016133:Polymerase Chain Reaction / Q000379:methods; D011451:Prostaglandin-Endoperoxide Synthases / Q000096:biosynthesis* / Q000235:genetics*; D012333:RNA, Messenger / Q000096:biosynthesis* / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D017386:Sequence Homology, Amino Acid; D014162:Transfection
D018076:DNA, Complementary; D008070:Lipopolysaccharides; D012333:RNA, Messenger; D002351:Carrageenan; D011451:Prostaglandin-Endoperoxide Synthases; D015232:Dinoprostone