Proteomics of the injured rat sciatic nerve reveals protein expression dynamics during regeneration

Connie R Jiménez; Floor J Stam; Ka Wan Li; Yvonne Gouwenberg; Martin P Hornshaw; Fred De Winter; J. Verhaagen; August B Smit

doi:10.1074/mcp.M400076-MCP200

Proteomics of the injured rat sciatic nerve reveals protein expression dynamics during regeneration

Connie R Jiménez, Floor J Stam, Ka Wan Li, Yvonne Gouwenberg, Martin P Hornshaw, Fred De Winter, J. Verhaagen, August B Smit

Netherlands Institute for Neuroscience (NIN)

Research output: Contribution to journal/periodical › Article › Scientific › peer-review

61 Citations (Scopus)

Abstract

Using proteomics, we investigated the temporal expression profiles of proteins in rat sciatic nerve after experimental crush. Extracts of sciatic nerves collected at 5, 10, and 35 days after injury were analyzed by two-dimensional gel electrophoresis and quantitative image analysis. Of the approximately 1,500 protein spots resolved on each gel, 121 showed significant regulation during at least one time point. Using cluster analysis, these proteins were grouped into two expression profiles of down-regulation and four of up-regulation. These profiles mainly reflected differences in cellular origins in addition to different functional roles. Mass spectrometric analysis identified 82 proteins pertaining to several functional classes, i.e. acute-phase proteins, antioxidant proteins, and proteins involved in protein synthesis/maturation/degradation, cytoskeletal (re)organization, and in lipid metabolism. Several proteins not previously implicated in nerve regeneration were identified, e.g. translationally controlled tumor protein, annexin A9/31, vitamin D-binding protein, alpha-crystallin B, alpha-synuclein, dimethylargininases, and reticulocalbin. Real-time PCR analysis of selected genes showed which were expressed in the nerve versus the dorsal root ganglion neurons. In conclusion, this study highlights the complexity and temporal aspect of the molecular process underlying nerve regeneration and points to the importance of glial and inflammatory determinants.

Original language	English
Pages (from-to)	120-32
Number of pages	13
Journal	Molecular and Cellular Proteomics
Volume	4
Issue number	2
DOIs	https://doi.org/10.1074/mcp.M400076-MCP200
Publication status	Published - Feb 2005

Keywords

Animals
Base Sequence
Cluster Analysis
Cytoskeleton
DNA Primers
DNA, Complementary
Electrophoresis, Gel, Two-Dimensional
Gene Expression Regulation
Image Processing, Computer-Assisted
Inflammation
Male
Mass Spectrometry
Molecular Sequence Data
Polymerase Chain Reaction
Proteomics
RNA
RNA, Messenger
Rats
Rats, Wistar
Regeneration
Sciatic Nerve
Silver Staining
Time Factors
Up-Regulation
Wound Healing
Journal Article

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10.1074/mcp.M400076-MCP200

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title = "Proteomics of the injured rat sciatic nerve reveals protein expression dynamics during regeneration",

abstract = "Using proteomics, we investigated the temporal expression profiles of proteins in rat sciatic nerve after experimental crush. Extracts of sciatic nerves collected at 5, 10, and 35 days after injury were analyzed by two-dimensional gel electrophoresis and quantitative image analysis. Of the approximately 1,500 protein spots resolved on each gel, 121 showed significant regulation during at least one time point. Using cluster analysis, these proteins were grouped into two expression profiles of down-regulation and four of up-regulation. These profiles mainly reflected differences in cellular origins in addition to different functional roles. Mass spectrometric analysis identified 82 proteins pertaining to several functional classes, i.e. acute-phase proteins, antioxidant proteins, and proteins involved in protein synthesis/maturation/degradation, cytoskeletal (re)organization, and in lipid metabolism. Several proteins not previously implicated in nerve regeneration were identified, e.g. translationally controlled tumor protein, annexin A9/31, vitamin D-binding protein, alpha-crystallin B, alpha-synuclein, dimethylargininases, and reticulocalbin. Real-time PCR analysis of selected genes showed which were expressed in the nerve versus the dorsal root ganglion neurons. In conclusion, this study highlights the complexity and temporal aspect of the molecular process underlying nerve regeneration and points to the importance of glial and inflammatory determinants.",

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author = "Jim{\'e}nez, {Connie R} and Stam, {Floor J} and Li, {Ka Wan} and Yvonne Gouwenberg and Hornshaw, {Martin P} and {De Winter}, Fred and J. Verhaagen and Smit, {August B}",

year = "2005",

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doi = "10.1074/mcp.M400076-MCP200",

language = "English",

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AU - Jiménez, Connie R

AU - Stam, Floor J

AU - Li, Ka Wan

AU - Gouwenberg, Yvonne

AU - Hornshaw, Martin P

AU - De Winter, Fred

AU - Verhaagen, J.

AU - Smit, August B

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AB - Using proteomics, we investigated the temporal expression profiles of proteins in rat sciatic nerve after experimental crush. Extracts of sciatic nerves collected at 5, 10, and 35 days after injury were analyzed by two-dimensional gel electrophoresis and quantitative image analysis. Of the approximately 1,500 protein spots resolved on each gel, 121 showed significant regulation during at least one time point. Using cluster analysis, these proteins were grouped into two expression profiles of down-regulation and four of up-regulation. These profiles mainly reflected differences in cellular origins in addition to different functional roles. Mass spectrometric analysis identified 82 proteins pertaining to several functional classes, i.e. acute-phase proteins, antioxidant proteins, and proteins involved in protein synthesis/maturation/degradation, cytoskeletal (re)organization, and in lipid metabolism. Several proteins not previously implicated in nerve regeneration were identified, e.g. translationally controlled tumor protein, annexin A9/31, vitamin D-binding protein, alpha-crystallin B, alpha-synuclein, dimethylargininases, and reticulocalbin. Real-time PCR analysis of selected genes showed which were expressed in the nerve versus the dorsal root ganglion neurons. In conclusion, this study highlights the complexity and temporal aspect of the molecular process underlying nerve regeneration and points to the importance of glial and inflammatory determinants.

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KW - Gene Expression Regulation

KW - Image Processing, Computer-Assisted

KW - Inflammation

KW - Male

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KW - Molecular Sequence Data

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KW - Proteomics

KW - RNA

KW - RNA, Messenger

KW - Rats

KW - Rats, Wistar

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KW - Silver Staining

KW - Time Factors

KW - Up-Regulation

KW - Wound Healing

KW - Journal Article

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DO - 10.1074/mcp.M400076-MCP200

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SN - 1535-9476

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SP - 120

EP - 132

JO - Molecular and Cellular Proteomics

JF - Molecular and Cellular Proteomics

IS - 2

ER -

Proteomics of the injured rat sciatic nerve reveals protein expression dynamics during regeneration

Abstract

Keywords

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