Determination of changes in mRNA expression in a rat model of neuropathic pain by Taqman quantitative RT-PCR.
The aim of this study was to develop a rapid and accurate high throughput method of screening multiple genes across a single sample set to detect changes in gene expression in the dorsal root ganglion (DRG) following partial sciatic nerve ligation in the rat. Using Taqman quantitative RT-PCR, we show that expression of a number of genes, including galanin, vasointestinal peptide and neuropeptide Y are rapidly increased 24 h post-operation in the DRGs on the ligated side only. Other genes tested, including vanilloid receptor-1, substance P, galanin receptor-2 and housekeeping genes did not alter. Analysis of the expression of ASIC4 showed a small difference in expression at 7 days post ligation. By applying a statistical method for analysis of multiple variables, partial least squares, we show that the expression change of ASIC4 was significantly altered on the ligated side even though the change was small. This method will allow us to rapidly identify changes in expression of candidate genes that may be involved in adaptive responses in the DRG due to nerve injury.
Macdonald
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R
Neurology CEDD, GlaxoSmithKline, New Frontiers Science Park, Essex CM19 5AW, Harlow, UK. rachel_macdonald-1@sbphrd.com
Bingham
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S
Bond
B C
BC
Parsons
A A
AA
Philpott
K L
KL
D062565:Acid Sensing Ion Channels; D000818:Animals; D018076:DNA, Complementary / Q000235:genetics; D019004:Galanin / Q000096:biosynthesis / Q000235:genetics; D005727:Ganglia, Spinal / Q000378:metabolism*; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D006358:Hot Temperature; D006930:Hyperalgesia / Q000235:genetics / Q000378:metabolism; D008026:Ligation; D008297:Male; D008565:Membrane Proteins; D009419:Nerve Tissue Proteins / Q000096:biosynthesis* / Q000235:genetics; D009437:Neuralgia / Q000235:genetics / Q000378:metabolism*; D009475:Neurons, Afferent / Q000378:metabolism*; D009478:Neuropeptide Y / Q000096:biosynthesis* / Q000235:genetics; D017288:Pain Threshold; D012333:RNA, Messenger / Q000096:biosynthesis*; D051381:Rats; D017208:Rats, Wistar; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000295:instrumentation; D012584:Sciatic Nerve / Q000293:injuries; D015222:Sodium Channels / Q000096:biosynthesis / Q000235:genetics; D019914:Taq Polymerase; D014660:Vasoactive Intestinal Peptide / Q000096:biosynthesis* / Q000235:genetics
C112231:ASIC3 protein, human; C568270:ASIC4 protein, human; D062565:Acid Sensing Ion Channels; D018076:DNA, Complementary; D008565:Membrane Proteins; D009419:Nerve Tissue Proteins; D009478:Neuropeptide Y; D012333:RNA, Messenger; D015222:Sodium Channels; D014660:Vasoactive Intestinal Peptide; D019004:Galanin; D019914:Taq Polymerase