Proteomics of the injured rat sciatic nerve reveals protein expression dynamics during regeneration.
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.
Jiménez
Connie R
CR
Department of Molecular and Cellular Neurobiology, Research Institute Neurosciences Amsterdam, Faculty of Earth and Life Sciences, Vrije Universiteit Amsterdam, 1081 HV Amsterdam, The Netherlands. connie.jimenez@falw.vu.nl
Stam
Floor J
FJ
Li
Ka Wan
KW
Gouwenberg
Yvonne
Y
Hornshaw
Martin P
MP
De Winter
Fred
F
Verhaagen
Joost
J
Smit
August B
AB
D000818:Animals; D001483:Base Sequence; D016000:Cluster Analysis; D003599:Cytoskeleton / Q000378:metabolism; D017931:DNA Primers / Q000737:chemistry; D018076:DNA, Complementary / Q000378:metabolism; D015180:Electrophoresis, Gel, Two-Dimensional; D005786:Gene Expression Regulation; D007091:Image Processing, Computer-Assisted; D007249:Inflammation; D008297:Male; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D040901:Proteomics / Q000379:methods*; D012313:RNA / Q000737:chemistry / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D012038:Regeneration; D012584:Sciatic Nerve / Q000293:injuries* / Q000502:physiology*; D016622:Silver Staining; D013997:Time Factors; D015854:Up-Regulation; D014945:Wound Healing
D017931:DNA Primers; D018076:DNA, Complementary; D012333:RNA, Messenger; D012313:RNA