on the proteolysis of blood_brain barrier and white matter components after cerebral ischemia.pdfVIP

Effects of Matrix Metalloproteinase-9 Gene Knock-Out on the Proteolysis of Blood–Brain Barrier and White Matter Components after Cerebral Ischemia Minoru Asahi,1 Xiaoying Wang,1 Tatsuro Mori,1 Toshihisa Sumii,1 Jae-Chang Jung,2 Michael A. Moskowitz,3 M. Elizabeth Fini,2 and Eng H. Lo1 1Neuroprotection Research Laboratory, Departments of Neurology and Radiology, Massachusetts General Hospital, and Program in Neuroscience, Harvard Medical School, Charlestown, Massachusetts 02129, 2Vision Research Laboratories, New England Eye Center, Tufts University, Boston, Massachusetts 02111, and 3Stroke and Neurovascular Regulation Laboratory, Massachusetts General Hospital and Harvard Medical School, Charlestown, Massachusetts 02129 Deleterious processes of extracellular proteolysis may contrib- ute to the progression of tissue damage after acute brain injury. We recently showed that matrix metalloproteinase-9 (MMP-9) knock-out mice were protected against ischemic and traumatic brain injury. In this study, we examined the mechanisms in- volved by focusing on relevant MMP-9 substrates in blood– brain barrier, matrix, and white matter. MMP-9 knock-out and wild-type mice were subjected to transient focal ischemia. MMP-9 levels increased after ischemia in wild-type brain, with expression primarily present in vascular endothelium. Western blots showed that the blood–brain barrier-associated protein and MMP-9 substrate zonae occludens-1 was degraded after ischemia, but this was reduced in knock-out mice. There were no detectable changes in another blood–brain barrier- associated protein, occludin. Correspondingly, blood–brain barrier disruption assessed via Evans Blue leakage was signif- icantly attenuated in MMP-9 knock-out mice compared with wild types. In white matter, ischemic degradation of the MMP-9 substrate myelin basic protein was significantly reduced in knock-out mice compared with wild types, whereas there was no degradation of other myelin proteins that are not MMP substr