title: A Model of Excitotoxic Brain Injury in Larval Zebrafish: Potential Application for High-Throughput Drug Evaluation to Treat Traumatic Brain Injury.
abstract: Traumatic brain injury (TBI) is a leading cause of death and morbidity with no effective therapeutic treatments for secondary injury. Preclinical drug evaluation in rodent models of TBI is a lengthy process. In this regard, the zebrafish has numerous advantages to address the technical and time-dependent obstacles associated with drug evaluation. We developed a reproducible brain injury using glutamate excitoxicity in zebrafish larvae, a known initiator of delayed cell death in TBI. Glutamate challenge resulted in dose-dependent lethality over an 84-h observation period. We report significant decrease in locomotion (p < 0.0001) and mean velocity (p < 0.001) with 10 μM glutamate application as measured through automated 96-well plate behavioral analysis. Application of the NMDA receptor antagonist MK-801 (400 nM) or the calpain inhibitor, MDL-28170 (20 μM), resulted in significant recovery of locomotor function. A secA5-YFP transgenic line was used to visualize the localization of cell death due to glutamate exposure in vivo using confocal fluorescence microscopy. Our results indicate that zebrafish larvae exhibit responses to excitotoxic injury and pharmacotherapeutic intervention with pathophysiological relevance to mammalian excitotoxic brain injury. This system has potential to be applied as a high-throughput drug screening model to quickly identify candidate lead compounds for further evaluation.
authors.affiliation: 1 Institute of Medical Sciences, University of Toronto , Toronto, Ontario, Canada .|2 Keenan Research Centre for Biomedical Science, Li Ka Shing Knowledge Institute , St. Michael's Hospital, Toronto, Ontario, Canada .
authors.forename: Victoria
authors.initials: V
authors.lastname: McCutcheon
authors.affiliation: 2 Keenan Research Centre for Biomedical Science, Li Ka Shing Knowledge Institute , St. Michael's Hospital, Toronto, Ontario, Canada .
authors.forename: Eugene
authors.initials: E
authors.lastname: Park
authors.affiliation: 2 Keenan Research Centre for Biomedical Science, Li Ka Shing Knowledge Institute , St. Michael's Hospital, Toronto, Ontario, Canada .
authors.forename: Elaine
authors.initials: E
authors.lastname: Liu
authors.affiliation: 2 Keenan Research Centre for Biomedical Science, Li Ka Shing Knowledge Institute , St. Michael's Hospital, Toronto, Ontario, Canada .
authors.forename: Youdong
authors.initials: Y
authors.lastname: Wang
authors.affiliation: 1 Institute of Medical Sciences, University of Toronto , Toronto, Ontario, Canada .|2 Keenan Research Centre for Biomedical Science, Li Ka Shing Knowledge Institute , St. Michael's Hospital, Toronto, Ontario, Canada .|3 Department of Medicine, University of Toronto , Toronto, Ontario, Canada .|4 Department of Physiology, University of Toronto , Toronto, Ontario, Canada .
authors.forename: Xiao-Yan
authors.initials: XY
authors.lastname: Wen
authors.affiliation: 1 Institute of Medical Sciences, University of Toronto , Toronto, Ontario, Canada .|2 Keenan Research Centre for Biomedical Science, Li Ka Shing Knowledge Institute , St. Michael's Hospital, Toronto, Ontario, Canada .|5 Department of Anesthesia, University of Toronto , Toronto, Ontario, Canada .|6 Department of Surgery, University of Toronto , Toronto, Ontario, Canada .
authors.forename: Andrew J
authors.initials: AJ
authors.lastname: Baker
mesh_terms: D000818:Animals; D001930:Brain Injuries / Q000188:drug therapy* / Q000473:pathology; D004151:Dipeptides / Q000494:pharmacology; D016291:Dizocilpine Maleate / Q000494:pharmacology; D057166:High-Throughput Screening Assays / Q000379:methods*; D007814:Larva; D009043:Motor Activity; D018696:Neuroprotective Agents / Q000494:pharmacology*; D015027:Zebrafish
chemical_list: D004151:Dipeptides; D018696:Neuroprotective Agents; D016291:Dizocilpine Maleate; C058076:calpain inhibitor III