Glutathione peroxidase activity modulates recovery in the injured immature brain†
Kyoko Tsuru-Aoyagi MD
Department of Neurological Surgery, University of California, San Francisco, San Francisco, CA
Search for more papers by this authorMatthew B. Potts MD
Department of Neurological Surgery, University of California, San Francisco, San Francisco, CA
Search for more papers by this authorAlpa Trivedi PhD
Department of Neurological Surgery, University of California, San Francisco, San Francisco, CA
Search for more papers by this authorTimothy Pfankuch BS
Department of Behavioral Neuroscience, Oregon National Primate Research Center, Oregon Health and Science University, Portland, OR
Search for more papers by this authorJacob Raber PhD
Department of Behavioral Neuroscience, Oregon National Primate Research Center, Oregon Health and Science University, Portland, OR
Department of Neurology, Oregon National Primate Research Center, Oregon Health and Science University, Portland, OR
Division of Neuroscience, Oregon National Primate Research Center, Oregon Health and Science University, Portland, OR
Search for more papers by this authorMichael Wendland PhD
Department of Radiology, University of California, San Francisco, San Francisco, CA
Search for more papers by this authorCatherine P. Claus BS
Department of Neurological Surgery, University of California, San Francisco, San Francisco, CA
Search for more papers by this authorSeong-Eun Koh MD
Department of Neurological Surgery, University of California, San Francisco, San Francisco, CA
Search for more papers by this authorDonna Ferriero MD
Department of Neurology, University of California, San Francisco, San Francisco, CA
Department of Pediatrics, University of California, San Francisco, San Francisco, CA
Search for more papers by this authorCorresponding Author
Linda J. Noble-Haeusslein PhD
Department of Neurological Surgery, University of California, San Francisco, San Francisco, CA
Department of Physical Therapy and Rehabilitation Science, University of California, San Francisco, San Francisco, CA
Departments of Neurological Surgery and Physical Therapy and Rehabilitation Science, University of California, San Francisco, 521 Parnassus Avenue, Room C-224, San Francisco, CA 94143-0520Search for more papers by this authorKyoko Tsuru-Aoyagi MD
Department of Neurological Surgery, University of California, San Francisco, San Francisco, CA
Search for more papers by this authorMatthew B. Potts MD
Department of Neurological Surgery, University of California, San Francisco, San Francisco, CA
Search for more papers by this authorAlpa Trivedi PhD
Department of Neurological Surgery, University of California, San Francisco, San Francisco, CA
Search for more papers by this authorTimothy Pfankuch BS
Department of Behavioral Neuroscience, Oregon National Primate Research Center, Oregon Health and Science University, Portland, OR
Search for more papers by this authorJacob Raber PhD
Department of Behavioral Neuroscience, Oregon National Primate Research Center, Oregon Health and Science University, Portland, OR
Department of Neurology, Oregon National Primate Research Center, Oregon Health and Science University, Portland, OR
Division of Neuroscience, Oregon National Primate Research Center, Oregon Health and Science University, Portland, OR
Search for more papers by this authorMichael Wendland PhD
Department of Radiology, University of California, San Francisco, San Francisco, CA
Search for more papers by this authorCatherine P. Claus BS
Department of Neurological Surgery, University of California, San Francisco, San Francisco, CA
Search for more papers by this authorSeong-Eun Koh MD
Department of Neurological Surgery, University of California, San Francisco, San Francisco, CA
Search for more papers by this authorDonna Ferriero MD
Department of Neurology, University of California, San Francisco, San Francisco, CA
Department of Pediatrics, University of California, San Francisco, San Francisco, CA
Search for more papers by this authorCorresponding Author
Linda J. Noble-Haeusslein PhD
Department of Neurological Surgery, University of California, San Francisco, San Francisco, CA
Department of Physical Therapy and Rehabilitation Science, University of California, San Francisco, San Francisco, CA
Departments of Neurological Surgery and Physical Therapy and Rehabilitation Science, University of California, San Francisco, 521 Parnassus Avenue, Room C-224, San Francisco, CA 94143-0520Search for more papers by this authorPotential conflict of interest: Nothing to report.
Abstract
Objective
Mice subjected to traumatic brain injury at postnatal day 21 show emerging cognitive deficits that coincide with hippocampal neuronal loss. Here we consider glutathione peroxidase (GPx) activity as a determinant of recovery in the injured immature brain.
Methods
Wild-type and transgenic (GPxTg) mice overexpressing GPx were subjected to traumatic brain injury or sham surgery at postnatal day 21. Animals were killed acutely (3 or 24 hours after injury) to assess oxidative stress and cell injury in the hippocampus or 4 months after injury after behavioral assessments.
Results
In the acutely injured brains, a reduction in oxidative stress markers including nitrotyrosine was seen in the injured GPxTg group relative to wild-type control mice. In contrast, cell injury, with marked vulnerability in the dentate gyrus, was apparent despite no differences between genotypes. Magnetic resonance imaging demonstrated an emerging cortical lesion during brain maturation that was also indistinguishable between injured genotypes. Stereological analyses of cortical volumes likewise confirmed no genotypic differences between injured groups. However, behavioral tests beginning 3 months after injury demonstrated improved spatial memory learning in the GPxTg group. Moreover, stereological analysis within hippocampal subregions demonstrated a significantly greater number of neurons within the dentate of the GPx group.
Interpretation
Our results implicate GPx in recovery of spatial memory after traumatic brain injury. This recovery may be attributed, in part, to a reduction in early oxidative stress and selective, long-term sparing of neurons in the dentate. Ann Neurol 2009;65:540–549
Supporting Information
Additional Supporting Information may be found in the online version of this article.
| Filename | Description |
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| ANA_21600_sm_SupFig1.tif35.2 MB | Supplemental Figure 1. TUNEL positive nuclei in the hippocampus at 24 hours postinjury. A–D) Representative photomicrographs of TUNEL staining, an indicator of irreversible cell injury. TUNEL positive cells are prominent in the granule cell layer of the dentate gyrus (arrows) in both genotypes. Scale bar, 100μm. E) The percentage of TUNEL-positive cells [(number of TUNEL-positive nuclei/number of Hoechst-positive nuclei) × 100%] was determined within 5 regions of the hippocampus for each genotype. The numbers of labeled nuclei are more prominent in caudal hippocampus relative to rostral. However, no differences in TUNEL labeling are noted between genotypes. |
| ANA_21600_sm_SupFig2.tif19 MB | Supplemental Figure 2. Temporal changes in the cortical lesion as assessed by magnetic resonance imaging. A) Representative T2WI and DWI from a WT injured animal. At 1 day postinjury, the site of injury exhibits relative high signal intensity on T2 images, suggesting vasogenic edema, and high signal intensity on DWI. Calculated ADCs are abnormally low for the injured region, suggesting cytotoxic edema was also present. At 7 days postinjury the lesion consists of both a hypointense region on T2 (arrows), consistent with aged clot, and hyperintense regions on T2 whose signal are almost nullified on DWI (not present in selected case) consistent with liquified tissue. At 14 and 24 days postinjury, the lesion consists of liquified tissue. B) Time course of signal increase in injured brain over 30 min after contrast administration at 1 day postinjury. This graph shows a relatively steady signal increase over time after contrast administration consistent with leakage of contrast material into the injured tissue. Minor but non-significant differences between WT and GPxTg groups can be ascribed to somewhat smaller injury among the GPxTg animals examined. |
| ANA_21600_sm_SupFig3.tif34.5 MB | Supplemental Figure 3. Exploratory activity and anxiety levels after TBI or sham surgery. Exploratory activity and anxiety levels were assessed in the open field. A) GPxTg animals spend more time in the center of the open field (# p<0.05), indicating lower levels of anxiety than WT mice. B) There is no difference in total distance moved, an indication of total activity levels. C, D) Within the GPxTg group, injured mice enter the center less (C, * p<0.05) and move less in the center (D, * p<0.05). |
| ANA_21600_sm_SupFig4.tif27.5 MB | Supplemental Figure 4. Sensorimotor learning after TBI or sham surgery. Sensorimotor function was assessed using the rotorod. A) All groups exhibit improvement with training. B) The GPxTg animals that received TBI show significantly less improvement in rotorod performance with training than GPxTg sham mice (difference in time between trial 9 and trial 1). * p<0.05 |
| ANA_21600_sm_SupFig5.tif34.4 MB | Supplemental Figure 5. Cortical and hippocampal lesion volumes at 4 months postinjury. A, B) Cresyl violet staining demonstrates how a cortical cavitation replaces the frontal and parietal gray matter and subcortical white matter. Scale bars, 10μm. C, D) Regional volumes of ipsilateral cortex and hippocampus, estimated by the Cavalieri method, are reduced after TBI. However, this reduction is similar between genotypes (unpaired T-tests; p=0.622 and P=0.512 for the cortex and hippocampus, respectively). |
| ANA_21600_sm_SupMethods.doc44.5 KB | Supplemental methods |
Please note: The publisher is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.
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