Abstract
Neuroprotection in acute stroke has not been successfully translated from animals to humans. Animal research on promising agents continues largely in rats and mice which are commonly available to researchers. However, controversies continue on the most suitable species to model the human situation. Generally, putative agents seem less effective in mice as compared with rats. We hypothesized that this may be due to inter-species differences in stroke response and that this might be manifest at a genetic level. Here we used whole-genome microarrays to examine the differential gene regulation in the ischemic penumbra of mice and rats at 2 and 6 h after permanent middle cerebral artery occlusion (pMCAO; Raw microarray CEL data files are available in the GEO database with an accession number GSE163654). Differentially expressed genes (adj. p ≤ 0.05) were organized by hierarchical clustering, correlation plots, Venn diagrams and pathway analyses in each species and at each time-point. Emphasis was placed on genes already known to be associated with stroke, including validation by RT-PCR. Gene expression patterns in the ischemic penumbra differed strikingly between the species at both 2 h and 6 h. Nearly 90% of significantly regulated genes and most pathways modulated by ischemia differed between mice and rats. These differences were evident globally, among stroke-associated genes, immediate early genes, genes implicated in stress response, inflammation, neuroprotection, ion channels, and signal transduction. The findings of this study may have significant implications for the choice of species for screening putative stroke therapies.
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All data in this paper are available to scientific communities upon reasonable request to the corresponding author.
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Acknowledgements
We thank Dr. Lu-Yang Wang and Dr. Michael Salter for their critical review.
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This work was supported by funds from the Canada Research Chairs program.
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MT, QJW, XS, designed the experiments. QJW, XS, LT and DM performed the experiments. QJW and MT wrote the manuscript.
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Dr. Michael Tymianski (M.T.) is a Canada Research Chair (Tier 1) in Translational Stroke Research, and the CEO of NoNO Inc., a biotechnology company developing nerinetide (also termed NA-1 or Tat-NR2B9c) for clinical use. Q.J.W., X.J.S., L.T. and D.M. have no competing interests.
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All procedures performed involving animals were approved by the University Health Network animal care committee, conformed to Canadian Council of Animal Care guidelines, and ARRIVE guidelines (Kilkenny et al. 2010).
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10571_2021_1138_MOESM1_ESM.tif
Supplementary Figure 1 Method of determination of sampling sites in the ischemic penumbras of rats or mice following pMCAO. a–c Staining of animal brains (rat brain shown here) using triphenyltetrazolium chloride (TTC) at 2 h (a), 6 h (b) and 24 h (c) post-MCAO, to determine the regions of the brain cortex that are viable (red) at 2 h and 6 h and then will go on to stroke infarction (white) at 24 h. The stroke infarcts at 6 h and 24 h are outlined by the black line. d–f Schematic brain sections showing the region of penumbra (pink region) at the time of sample collection at 2 h (d) and 6 h (e). Stroke infarction is shown as the gray shaded area at 6 h (e) and 24 h (h). The cortical brain samples collected at 2 h and 6 h post-MCAO for microarray analysis are marked by the circles in d–e. Supplementary file1 (TIF 791 kb)
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Wu, Q.J., Sun, X., Teves, L. et al. Mice and Rats Exhibit Striking Inter-species Differences in Gene Response to Acute Stroke. Cell Mol Neurobiol 42, 2773–2789 (2022). https://doi.org/10.1007/s10571-021-01138-8
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DOI: https://doi.org/10.1007/s10571-021-01138-8