Real-time quantitative PCR analysis of temporal-spatial alterations in gene expression after spinal cord contusion.
Rat spinal cord contusion injury models the histopathology associated with much clinical spinal cord injury (SCI). Studies on altered gene expression after SCI in these models may identify therapeutic targets for reducing secondary injury after the initial trauma and/or enhancing recovery processes. However, complex spatial and temporal alterations after injury could complicate interpretation of changes in gene expression. To test this hypothesis, we selected six genes and studied their temporal and spatial patterns of expression at 1 h, 1, 3 and 7 days after a standardized spinal cord contusion produced by a weight drop device (10 g x 25 mm at T8). Real-time RT-PCR using TaqMan probes was employed to quantify mRNA for proteolipid protein, glyceraldehyde-3-phosphate dehydrogenase, glial fibrillary acidic protein, nestin, and the GluR2 and NR1 subunits of glutamate receptors. We found widely different temporal and spatial patterns of altered gene expression after SCI, including instances of opposing up- and down-regulation at different locations in tissue immediately adjacent to the injury site. We conclude that greater use of the reliable and extremely sensitive technique of quantitative real-time PCR for regional tissue analysis is important for understanding the altered gene expression that occurs after CNS trauma.
Wu
Xiaofang
X
Department of Neuroscience, Georgetown University Medical Center, Washington, District of Columbia 20057, USA.
Yoo
Soonmoon
S
Wrathall
Jean R
JR
D000818:Animals; D015152:Blotting, Northern; D001921:Brain / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000378:metabolism; D005260:Female; D005786:Gene Expression Regulation / Q000502:physiology*; D005904:Glial Fibrillary Acidic Protein / Q000378:metabolism; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000378:metabolism; D007381:Intermediate Filament Proteins / Q000378:metabolism; D018991:Myelin Proteolipid Protein; D009419:Nerve Tissue Proteins / Q000378:metabolism; D064231:Nestin; D050811:Octamer Transcription Factor-1; D010446:Peptide Fragments / Q000378:metabolism; D018272:Porins / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D018091:Receptors, AMPA / Q000378:metabolism; D016194:Receptors, N-Methyl-D-Aspartate / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013119:Spinal Cord Injuries / Q000235:genetics / Q000378:metabolism* / Q000503:physiopathology; D013997:Time Factors; D014157:Transcription Factors / Q000378:metabolism
D004268:DNA-Binding Proteins; D005904:Glial Fibrillary Acidic Protein; D007381:Intermediate Filament Proteins; D018991:Myelin Proteolipid Protein; C409466:NR1 NMDA receptor; D009419:Nerve Tissue Proteins; C577486:Nes protein, rat; D064231:Nestin; D050811:Octamer Transcription Factor-1; D010446:Peptide Fragments; D018272:Porins; C494168:Pou2f1 protein, rat; D012333:RNA, Messenger; D018091:Receptors, AMPA; D016194:Receptors, N-Methyl-D-Aspartate; D014157:Transcription Factors; C068142:glyceraldehyde 3-phosphate dehydrogenase (304-313); D005987:Glyceraldehyde-3-Phosphate Dehydrogenases; C104722:glutamate receptor ionotropic, AMPA 2