Iadecola C, Niwa K, Nogawa S, Zhao X, Nagayama M, Araki E, Morham S, Ross ME.
Reduced susceptibility to ischemic brain injury and N-methyl-D-aspartate-mediated neurotoxicity in cyclooxygenase-2-deficient mice.Proc Natl Acad Sci U S A.
2001 Jan 30;98(3):1294-9.
REAGENTS/MATERIAL: COX-2-null mice obtained from University of Minnesota.
Mice (SV129 X C57BL/6J) were backcrossed to C57BL/6J mice five or six times.
First-strand cDNA synthesis was carried out with Moloney murine leukemia virus reverse transcriptase (New England Biolabs)
COX-2 primers: Forward 5'-CCAGATGCTATCTTTGGGGAGAC-3'
COX-1 primers: Forward 5'-GAATACCGAAAGAGGTTTGGCTTG-3'
PBD primers: Forward 5'-GCCACCACAGTCTCGGTCTGTATGCGAGC-3'
Deletion construct miss an internal 79 nucleotide fragment from the endogenous COX-2 message
restriction enzyme, Sau3AI (New England Biolabs)
T4 DNA ligase (New England Biolabs)
FUTURE DIRECTION: Further elucidation on the mechanisms by which COX-2 contributes to neurotoxicity, and the role of PGE2, a reaction product of COX-2 in neurotoxicity. COX-2 inhibitors, a potential pharmacological target for the treatment of ischemic stroke.