title: Gene expression analysis to identify molecular correlates of pre- and post-conditioning derived neuroprotection.
abstract: Mild ischaemic exposures before or after severe injurious ischaemia that elicit neuroprotective responses are referred to as preconditioning and post-conditioning. The corresponding molecular mechanisms of neuroprotection are not completely understood. Identification of the genes and associated pathways of corresponding neuroprotection would provide insight into neuronal survival, potential therapeutic approaches and assessments of therapies for stroke. The objectives of this study were to use global gene expression approach to infer the molecular mechanisms in pre- and post-conditioning-derived neuroprotection in cortical neurons following oxygen and glucose deprivation (OGD) in vitro and then to apply these findings to predict corresponding functional pathways. To this end, microarray analysis was applied to rat cortical neurons with or without the pre- and post-conditioning treatments at 3-h post-reperfusion, and differentially expressed transcripts were subjected to statistical, hierarchical clustering and pathway analyses. The expression patterns of 3,431 genes altered under all conditions of ischaemia (with and without pre- or post-conditioning). We identified 1,595 genes that were commonly regulated within both the pre- and post-conditioning treatments. Cluster analysis revealed that transcription profiles clustered tightly within controls, non-conditioned OGD and neuroprotected groups. Two clusters defining neuroprotective conditions associated with up- and downregulated genes were evident. The five most upregulated genes within the neuroprotective clusters were Tagln, Nes, Ptrf, Vim and Adamts9, and the five most downregulated genes were Slc7a3, Bex1, Brunol4, Nrxn3 and Cpne4. Pathway analysis revealed that the intracellular and second messenger signalling pathways in addition to cell death were predominantly associated with downregulated pre- and post-conditioning associated genes, suggesting that modulation of cell death and signal transduction pathways plays a role in the neuroprotection. A high degree of similarity in the pathways associated with the differentially expressed genes in the pre- and post-conditioning treatments suggests that similar molecular mechanisms may mediate their neuroprotective effects.
authors.affiliation: Genomics Laboratories, Biologics and Genetic Therapies Directorate, Health Canada, 251 Sir Frederick Banting Driveway, Ottawa, ON, K1A 0K9, Canada. Shiv.Prasad@hc-sc.gc.ca
authors.forename: Shiv S
authors.initials: SS
authors.lastname: Prasad
authors.forename: Marsha
authors.initials: M
authors.lastname: Russell
authors.forename: Margeryta
authors.initials: M
authors.lastname: Nowakowska
authors.forename: Andrew
authors.initials: A
authors.lastname: Williams
authors.forename: Carole
authors.initials: C
authors.lastname: Yauk
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000502:physiology; D019610:Cytoprotection / Q000235:genetics*; D005260:Female; D005786:Gene Expression Regulation / Q000502:physiology*; D019194:Ischemic Preconditioning / Q000379:methods*; D009474:Neurons / Q000166:cytology* / Q000502:physiology*; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods; D051381:Rats; D017208:Rats, Wistar; D059467:Transcriptome