Differential proteomic analysis of acute contusive spinal cord injury in rats using iTRAQ reagent labeling and LC-MS/MS

Anshu Chen; Shixin Sun; Rangaswamyrao Ravikumar; Nishant P. Visavadiya; Joe E. Springer

doi:10.1007/s11064-013-1132-y

Differential proteomic analysis of acute contusive spinal cord injury in rats using iTRAQ reagent labeling and LC-MS/MS

Anshu Chen, Shixin Sun, Rangaswamyrao Ravikumar, Nishant P. Visavadiya, Joe E. Springer

Research output: Contribution to journal › Article › peer-review

9 Scopus citations

Abstract

In this experimental study, differential labeling with isobaric tags for relative and absolute quantitation (iTRAQ) reagents followed by liquid chromatography (LC) and tandem mass spectrometry (MS/MS) proteomic approach was used to investigate differences in the proteome of rat spinal cord at 24 h following a moderate contusion injury. Spinal cord protein samples from the injury epicenter (or from sham controls) were trypsinized and differentially labeled with iTRAQ isotopic reagents. The differentially labeled samples were then combined into one sample mixture, separated by LC, and analyzed using MS/MS. Proteins were quantified by comparing the peak areas of iTRAQ reporter fragment ions in MS/MS spectra. The outcome of this analysis revealed that proteins involved in ubiquitination, endocytosis and exocytosis, energy metabolism, inflammatory response, oxidative stress, cytoskeletal disruption, and vascular damage were significantly altered at 24 h following spinal cord injury (SCI). This study demonstrates the utility of the iTRAQ method in proteomic studies and provides further insights into secondary events that occur during acute times following SCI.

Original language	English
Pages (from-to)	2247-2255
Number of pages	9
Journal	Neurochemical Research
Volume	38
Issue number	11
DOIs	https://doi.org/10.1007/s11064-013-1132-y
State	Published - Nov 2013

Bibliographical note

Funding Information:
Acknowledgments The authors gratefully acknowledge Dr. Melanie L. McEwen for technical support. This work was supported by the Craig H. Neilsen Foundation, the Morton Cure Paralysis Foundation, and an endowment from Cardinal Hill Rehabilitation Hospital.

Keywords

Contusion
LC/MS-MS analysis
Proteome
Spinal cord injury
iTRAQ

ASJC Scopus subject areas

Biochemistry
Cellular and Molecular Neuroscience

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title = "Differential proteomic analysis of acute contusive spinal cord injury in rats using iTRAQ reagent labeling and LC-MS/MS",

abstract = "In this experimental study, differential labeling with isobaric tags for relative and absolute quantitation (iTRAQ) reagents followed by liquid chromatography (LC) and tandem mass spectrometry (MS/MS) proteomic approach was used to investigate differences in the proteome of rat spinal cord at 24 h following a moderate contusion injury. Spinal cord protein samples from the injury epicenter (or from sham controls) were trypsinized and differentially labeled with iTRAQ isotopic reagents. The differentially labeled samples were then combined into one sample mixture, separated by LC, and analyzed using MS/MS. Proteins were quantified by comparing the peak areas of iTRAQ reporter fragment ions in MS/MS spectra. The outcome of this analysis revealed that proteins involved in ubiquitination, endocytosis and exocytosis, energy metabolism, inflammatory response, oxidative stress, cytoskeletal disruption, and vascular damage were significantly altered at 24 h following spinal cord injury (SCI). This study demonstrates the utility of the iTRAQ method in proteomic studies and provides further insights into secondary events that occur during acute times following SCI.",

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author = "Anshu Chen and Shixin Sun and Rangaswamyrao Ravikumar and Visavadiya, {Nishant P.} and Springer, {Joe E.}",

note = "Funding Information: Acknowledgments The authors gratefully acknowledge Dr. Melanie L. McEwen for technical support. This work was supported by the Craig H. Neilsen Foundation, the Morton Cure Paralysis Foundation, and an endowment from Cardinal Hill Rehabilitation Hospital.",

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AU - Visavadiya, Nishant P.

AU - Springer, Joe E.

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N2 - In this experimental study, differential labeling with isobaric tags for relative and absolute quantitation (iTRAQ) reagents followed by liquid chromatography (LC) and tandem mass spectrometry (MS/MS) proteomic approach was used to investigate differences in the proteome of rat spinal cord at 24 h following a moderate contusion injury. Spinal cord protein samples from the injury epicenter (or from sham controls) were trypsinized and differentially labeled with iTRAQ isotopic reagents. The differentially labeled samples were then combined into one sample mixture, separated by LC, and analyzed using MS/MS. Proteins were quantified by comparing the peak areas of iTRAQ reporter fragment ions in MS/MS spectra. The outcome of this analysis revealed that proteins involved in ubiquitination, endocytosis and exocytosis, energy metabolism, inflammatory response, oxidative stress, cytoskeletal disruption, and vascular damage were significantly altered at 24 h following spinal cord injury (SCI). This study demonstrates the utility of the iTRAQ method in proteomic studies and provides further insights into secondary events that occur during acute times following SCI.

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Differential proteomic analysis of acute contusive spinal cord injury in rats using iTRAQ reagent labeling and LC-MS/MS

Abstract

Bibliographical note

Keywords

ASJC Scopus subject areas

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