Intracellular-specific colocalization of prostaglandin E2 synthases and cyclooxygenases in the brain.
Prostaglandin E2 (PGE2) is the major primary prostaglandin generated by brain cells. However, the coordination and intracellular localization of the cyclooxygenases (COXs) and prostaglandin E synthases (PGESs) that convert arachidonic acid to PGE2 in brain tissue are not known. We aimed to determine whether microsomal and cytosolic PGES (mPGES-1 and cPGES) colocalize and coordinate activity with either COX-1 or COX-2 in brain tissue, particularly during development. Importantly, we found that cytosolic PGES also associates with microsomes (cPGES-m) from the cerebrum and cerebral vasculature of the pig and rat as well as microsomes from various cell lines; this seemed dependent on the carboxyl terminal 35-amino acid domain and a cysteine residue (C58) of cPGES. In microsomal membranes from the postnatal brain and cerebral microvessels of mature animals, cPGES-m colocalized with both COX-1 and COX-2, whereas mPGES-1 was undetectable in these microsomes. Accordingly, in this cell compartment, cPGES could coordinate its activity with COX-2 and COX-1 (partly inhibited by NS398); albeit in microsomes of the brain microvasculature from newborns, mPGES-1 was also present. In contrast, in nuclei of brain parenchymal and endothelial cells, mPGES-1 and cPGES colocalized exclusively with COX-2 (determined by immunoblotting and immunohistochemistry); these PGESs contributed to conversion of PGH2 into PGE2. Hence, contrary to a previously proposed model of exclusive COX-2/mPGES-1 coordination, COX-2 can coordinate with mPGES-1 and/or cPGES in the brain, depending on the cell compartment and the age group.
Deptartment of Pediatrics, Research Center of Hôpital Sainte-Justine, 3175, Côte Sainte-Catherine, Montréal, Québec, Canada H3T 1C5.
Alejandro
A
Vazquez-Tello
Li
L
Fan
Xin
X
Hou
Jean-Sébastien
JS
Joyal
Joseph A
JA
Mancini
Christiane
C
Quiniou
Ronald I
RI
Clyman
Fernand
F
Gobeil
Daya R
DR
Varma
Sylvain
S
Chemtob
D000818:Animals; D000831:Animals, Newborn; D015153:Blotting, Western; D001921:Brain / Q000166:cytology / Q000201:enzymology*; D002196:Capillaries / Q000166:cytology / Q000201:enzymology; D002478:Cells, Cultured; D051545:Cyclooxygenase 1; D051546:Cyclooxygenase 2; D015232:Dinoprostone / Q000096:biosynthesis*; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D015971:Gene Expression Regulation, Enzymologic / Q000235:genetics / Q000502:physiology*; D007527:Isoenzymes / Q000378:metabolism; D008297:Male; D008565:Membrane Proteins; D018613:Microscopy, Confocal; D008861:Microsomes / Q000201:enzymology / Q000378:metabolism; D010957:Plasmids / Q000235:genetics; D011247:Pregnancy; D011451:Prostaglandin-Endoperoxide Synthases / Q000378:metabolism*; D051381:Rats; D017208:Rats, Wistar; D013552:Swine; D014162:Transfection
D007527:Isoenzymes; D008565:Membrane Proteins; D051545:Cyclooxygenase 1; D051546:Cyclooxygenase 2; D011451:Prostaglandin-Endoperoxide Synthases; C496513:Ptgs1 protein, rat; D015232:Dinoprostone