Effect of ischaemic preconditioning on genomic response to cerebral ischaemia: similarity to neuroprotective strategies in hibernation and hypoxia-tolerant states.
BACKGROUND
Molecular mechanisms of neuroprotection that lead to ischaemic tolerance are incompletely understood. Identification of genes involved in the process would provide insight into cell survival and therapeutic approaches for stroke. We developed a mouse model of neuroprotection in stroke and did gene expression profiling to identify potential neuroprotective genes and their associated pathways.


METHODS
Eight mice per condition were subjected to occlusion of the middle cerebral artery for 15 min (preconditioning), 60 min (injurious ischaemia), or preconditioning followed 72 h later by injurious ischaemia. RNA was extracted from the cortical regions of the ischaemic and non-ischaemic hemispheres. Three pools per condition were generated, and RNA was hybridised to oligonucleotide microarrays for comparison of ischaemic and non-ischaemic hemispheres. Real-time PCR and western blots were used to validate results. Follow-up experiments were done to address the biological relevance of findings.


RESULTS
Microarray analysis revealed changes in gene expression with little overlap among the conditions of injurious ischaemia, ischaemic preconditioning, or both. Injurious ischaemia induced upregulation of gene expression; 49 (86%) of 57 genes regulated showed increased expression in the ischaemic hemisphere. By contrast, preconditioning followed by injurious ischaemia resulted in pronounced downregulation; 47 (77%) of 61 regulated genes showed lower expression. Preconditioning resulted in transcriptional changes involved in suppression of metabolic pathways and immune responses, reduction of ion-channel activity, and decreased blood coagulation.


CONCLUSIONS
Preconditioning reprogrammes the response to ischaemic injury. Similar changes reported by others support an evolutionarily conserved endogenous response to decreased blood flow and oxygen limitation such as seen during hibernation.
Stenzel-Poore
Mary P
MP
Department of Molecular Microbiology and Immunology, Oregon Health and Science University, Portland, OR 97239, USA. poorem@ohsu.edu
Stevens
Susan L
SL
Xiong
Zhigang
Z
Lessov
Nikola S
NS
Harrington
Christina A
CA
Mori
Motomi
M
Meller
Robert
R
Rosenzweig
Holly L
HL
Tobar
Eric
E
Shaw
Tatyana E
TE
Chu
Xiangping
X
Simon
Roger P
RP
D000818:Animals; D001760:Bleeding Time; D015153:Blotting, Western; D001921:Brain / Q000098:blood supply / Q000473:pathology; D002545:Brain Ischemia / Q000235:genetics* / Q000503:physiopathology / Q000517:prevention & control*; D002470:Cell Survival / Q000235:genetics / Q000502:physiology; D002478:Cells, Cultured; D002560:Cerebrovascular Circulation / Q000235:genetics / Q000502:physiology*; D004195:Disease Models, Animal; D015870:Gene Expression / Q000502:physiology*; D006605:Hibernation / Q000235:genetics / Q000502:physiology*; D006801:Humans; D066298:In Vitro Techniques; D019194:Ischemic Preconditioning / Q000379:methods* / Q000592:standards; D051379:Mice; D018696:Neuroprotective Agents / Q000627:therapeutic use; D020411:Oligonucleotide Array Sequence Analysis; D016133:Polymerase Chain Reaction; D051381:Rats; D017207:Rats, Sprague-Dawley; D020521:Stroke / Q000473:pathology / Q000503:physiopathology* / Q000628:therapy
D018696:Neuroprotective Agents