Identification of injury specific proteins in a cell culture model of traumatic brain injury.

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...

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Main Authors: Camilla Lööv, Ganna Shevchenko, Aishwarya Geeyarpuram Nadadhur, Fredrik Clausen, Lars Hillered, Magnus Wetterhall, Anna Erlandsson
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2013-01-01
Series:PLoS ONE
Online Access:http://europepmc.org/articles/PMC3567017?pdf=render
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spelling doaj-f3f3d58c0131473aa7742f9e48d1a9dd2020-11-25T02:42:46ZengPublic Library of Science (PLoS)PLoS ONE1932-62032013-01-0182e5598310.1371/journal.pone.0055983Identification of injury specific proteins in a cell culture model of traumatic brain injury.Camilla LöövGanna ShevchenkoAishwarya Geeyarpuram NadadhurFredrik ClausenLars HilleredMagnus WetterhallAnna ErlandssonThe 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.http://europepmc.org/articles/PMC3567017?pdf=render
collection DOAJ
language English
format Article
sources DOAJ
author Camilla Lööv
Ganna Shevchenko
Aishwarya Geeyarpuram Nadadhur
Fredrik Clausen
Lars Hillered
Magnus Wetterhall
Anna Erlandsson
spellingShingle Camilla Lööv
Ganna Shevchenko
Aishwarya Geeyarpuram Nadadhur
Fredrik Clausen
Lars Hillered
Magnus Wetterhall
Anna Erlandsson
Identification of injury specific proteins in a cell culture model of traumatic brain injury.
PLoS ONE
author_facet Camilla Lööv
Ganna Shevchenko
Aishwarya Geeyarpuram Nadadhur
Fredrik Clausen
Lars Hillered
Magnus Wetterhall
Anna Erlandsson
author_sort Camilla Lööv
title Identification of injury specific proteins in a cell culture model of traumatic brain injury.
title_short Identification of injury specific proteins in a cell culture model of traumatic brain injury.
title_full Identification of injury specific proteins in a cell culture model of traumatic brain injury.
title_fullStr Identification of injury specific proteins in a cell culture model of traumatic brain injury.
title_full_unstemmed Identification of injury specific proteins in a cell culture model of traumatic brain injury.
title_sort identification of injury specific proteins in a cell culture model of traumatic brain injury.
publisher Public Library of Science (PLoS)
series PLoS ONE
issn 1932-6203
publishDate 2013-01-01
description 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.
url http://europepmc.org/articles/PMC3567017?pdf=render
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