title: Identification of injury specific proteins in a cell culture model of traumatic brain injury.
abstract: The complicated secondary molecular and cellular mechanisms following traumatic brain injury (TBI) are still not fully understood. In the present study, we have used mass spectrometry to identify injury specific proteins in an in vitro model of TBI. A standardized injury was induced by scalpel cuts through a mixed cell culture of astrocytes, oligodendrocytes and neurons. Twenty-four hours after the injury, cell culture medium and whole-cell fractions were collected for analysis. We found 53 medium proteins and 46 cell fraction proteins that were specifically expressed after injury and the known function of these proteins was elucidated by an extensive literature survey. By using time-lapse microscopy and immunostainings we could link a large proportion of the proteins to specific cellular processes that occur in response to trauma; including cell death, proliferation, lamellipodia formation, axonal regeneration, actin remodeling, migration and inflammation. A high percentage of the proteins uniquely expressed in the medium after injury were actin-related proteins, which normally are situated intracellularly. We show that two of these, ezrin and moesin, are expressed by astrocytes both in the cell culture model and in mouse brain subjected to experimental TBI. Interestingly, we found many inflammation-related proteins, despite the fact that cells were present in the culture. This study contributes with important knowledge about the cellular responses after trauma and identifies several potential cell-specific biomarkers.
authors: Department of Neuroscience, Uppsala University, Uppsala, Sweden.
authors: Camilla
authors: C
authors: Lööv
authors: Ganna
authors: G
authors: Shevchenko
authors: Aishwarya
authors: A
authors: Geeyarpuram Nadadhur
authors: Fredrik
authors: F
authors: Clausen
authors: Lars
authors: L
authors: Hillered
authors: Magnus
authors: M
authors: Wetterhall
authors: Anna
authors: A
authors: Erlandsson
mesh_terms: D000199:Actins / Q000378:metabolism; D000818:Animals; D001253:Astrocytes / Q000378:metabolism; D001369:Axons / Q000502:physiology; D001930:Brain Injuries / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D018929:Cell Culture Techniques; D016923:Cell Death; D002465:Cell Movement; D049109:Cell Proliferation; D002470:Cell Survival; D003598:Cytoskeletal Proteins / Q000378:metabolism; D013058:Mass Spectrometry; D051379:Mice; D008840:Microfilament Proteins / Q000378:metabolism; D009416:Nerve Regeneration; D009474:Neurons / Q000378:metabolism; D009836:Oligodendroglia / Q000378:metabolism; D009928:Organ Specificity; D010766:Phosphorylation; D020543:Proteome; D040901:Proteomics; D000099149:Ezrin
chemical_list: D000199:Actins; D003598:Cytoskeletal Proteins; D008840:Microfilament Proteins; D020543:Proteome; D000099149:Ezrin; C071058:moesin