Delayed neuromotor recovery and increased memory acquisition dysfunction following experimental brain trauma in mice lacking the DNA repair gene XPA

Laboratory investigation

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Object

This study investigates the outcome after traumatic brain injury (TBI) in mice lacking the essential DNA repair gene xeroderma pigmentosum group A (XPA). As damage to DNA has been implicated in neuronal cell death in various models, the authors sought to elucidate whether the absence of an essential DNA repair factor would affect the outcome of TBI in an experimental setting.

Methods

Thirty-seven adult mice of either wild-type (n = 18) or XPA-deficient (“knock-out” [n = 19]) genotype were subjected to controlled cortical impact experimental brain trauma, which produced a focal brain injury. Sham-injured mice of both genotypes were used as controls (9 in each group). The mice were subjected to neurobehavoral tests evaluating learning/acquisition (Morris water maze) and motor dysfunction (Rotarod and composite neuroscore test), pre- and postinjury up to 4 weeks. The mice were killed after 1 or 4 weeks, and cortical lesion volume, as well as hippocampal and thalamic cell loss, was evaluated. Hippocampal staining with doublecortin antibody was used to evaluate neurogenesis after the insult.

Results

Brain-injured XPA−/− mice exhibited delayed recovery from impairment in neurological motor function, as well as pronounced cognitive dysfunction in a spatial learning task (Morris water maze), compared with injured XPA+/+ mice (p < 0.05). No differences in cortical lesion volume, hippocampal damage, or thalamic cell loss were detected between XPA+/+ and XPA−/− mice after brain injury. Also, no difference in the number of cells stained with doublecortin in the hippocampus was detected.

Conclusions

The authors' results suggest that lack of the DNA repair factor XPA may delay neurobehavioral recovery after TBI, although they do not support the notion that this DNA repair deficiency results in increased cell or tissue death in the posttraumatic brain.

Abbreviations used in this paper:CCI = controlled cortical impact; MWM = Morris water maze; NER = nucleotide excision repair pathway; ROS = reactive oxygen species; TBI = traumatic brain injury; XP = xeroderma pigmentosum; XPA = XP complementation group A.

Article Information

Current affiliation for Dr. Mattiasson: Lund University and Skåne University Hospital, Lund, Sweden.

Address correspondence to: Gregor Tomasevic, M.D., Ph.D., Laboratory for Experimental Brain Research, Wallenberg Neuroscience Center, BMC A13, SE-221 84 Lund, Sweden. email: gregor.tomasevic@med.lu.se.

Please include this information when citing this paper: published online March 30, 2012; DOI: 10.3171/2012.2.JNS11888.

© AANS, except where prohibited by US copyright law.

Headings

Figures

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    Performance of XPA+/+ and XPA−/− mice in the composite neuroscore 48 hours to 4 weeks after CCI brain trauma. Brain-injured (INJ) mice of both genotypes achieved significantly lower scores than sham-operated (SH) mice at 48 hours and 1 week posttrauma (Kruskal-Wallis nonparametric test followed by Mann-Whitney test for comparison between individual groups). This deficiency persisted up to 2 weeks in brain-injured XPA−/− mice but only up to 1 week in XPA+/+ mice. No significant differences between brain-injured or sham-operated XPA+/+ and XPA−/− mice were found at any time point. Bar heights represent median scores of each group. Error bars represent the 25th percentiles. KO = knock out; WT = wild type.

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    Performance of XPA+/+ and XPA−/− mice in the Rotarod test (“fast” paradigm) 48 hours to 4 weeks after CCI brain trauma. Brain-injured mice of both genotypes exhibited significantly longer latencies than sham-operated mice at 48 hours posttrauma (Kruskal-Wallis nonparametric test followed by Mann-Whitney test for comparison between individual groups). This deficiency persisted at 1 week in brain-injured XPA−/− mice only. No significant differences between brain-injured or sham-operated XPA+/+ and XPA−/− mice were found at any time point. Bar heights represent the mean latency in percentage of baseline performance of each group. Error bars represent SDs.

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    Performance of XPA+/+ and XPA−/− mice in the MWM spatial memory (retention) paradigm 1 week after CCI brain trauma. The MWM score reflects latency to find the platform site as well as swim speed and time spent in the different zones surrounding the platform site. Brain-injured mice of both genotypes achieved significantly lower MWM scores than sham-operated mice at this time point (Kruskal-Wallis nonparametric test followed by Mann-Whitney test for comparison between individual groups). Also, sham-operated XPA−/− mice performed worse than sham-operated XPA+/+ mice. No significant difference between brain-injured XPA+/+ and XPA−/− mice was found at this time point. Bar heights represent median scores of each group. Error bars represent 25th percentiles.

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    Performance of XPA+/+ and XPA−/− mice in the MWM spatial learning (acquisition) paradigm 4 weeks after CCI brain trauma. Results are expressed as the sum of latencies to find the platform in the second day of learning (10 trials). Brain-injured XPA−/− mice performed significantly worse than sham-operated mice of the same genotype at this time point (1-way ANOVA followed by the Scheffé post hoc test for comparisons between individual groups). Brain-injured XPA−/− mice also performed worse than both injured and sham-operated XPA+/+ mice. No difference between any of the other groups was found in this test. Bar heights represent mean sum of latencies of each group. Error bars represent SDs.

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    Lesion volumes of brain-injured XPA+/+ and XPA−/− mice in the cortex ipsilateral to the site of injury 1 and 4 weeks after CCI brain trauma. No significant difference was found between injured XPA+/+ and XPA−/− mice at either time point. Also, no difference in lesion volumes was detected between the 2 time points in either genotype (unpaired Student t-test followed by Bonferroni correction for multiple comparisons). Bar heights represent mean cortical lesion volume in mm3. Error bars represent SDs.

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    Hippocampal damage scores of brain-injured and sham-operated XPA+/+ and XPA−/− mice in the CA1, CA3, and dentate gyrus (DG) regions 1 week after CCI brain trauma. Brain-injured XPA+/+ and XPA−/− mice exhibited significant damage in the CA1, CA3, and dentate gyrus hippocampal regions at this time point (Kruskal-Wallis nonparametric test followed by Mann-Whitney test for comparisons between individual groups). No difference was found between brain-injured or sham-operated XPA+/+ and XPA−/− mice in any of the regions at this time point. Bar heights represent median scores for each group. Error bars represent 25th percentiles.

  • View in gallery

    Hippocampal damage scores of brain-injured and sham-operated XPA+/+ and XPA−/− mice in the CA1, CA3, and dentate gyrus (DG) regions 4 weeks after CCI brain trauma. No significant damage was detected in any of the regions of XPA+/+ or XPA−/− mice at this time point (Kruskal-Wallis test). Also, no difference was found between the 2 genotypes after brain injury or sham surgery. Bar heights represent median scores for each group. Error bars represent 25th percentiles.

  • View in gallery

    Cell count in the ipsilateral thalamus of brain-injured and sham-operated XPA+/+ and XPA−/− mice 1 and 4 weeks after CCI brain injury. Significantly lower numbers of viable cells were detected in injured XPA+/+ and XPA−/− mice compared with sham-operated mice 1 week after brain injury (1-way ANOVA followed by Scheffé post hoc test). Also, significantly lower numbers of viable cells were detected in the ipsilateral thalamus of injured XPA+/+ mice at 4 weeks of recovery compared with sham-operated mice of the same genotype. No difference between brain-injured XPA+/+ and XPA−/− mice, or sham-operated mice of the 2 genotypes was detected at any of the time points. Bar heights represent the mean cell counts for each group. Error bars represent SDs.

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