ArticleMolecular pain2014
Validation of four reference genes for quantitative mRNA expression studies in a rat model of inflammatory injury.
Article in Molecular pain, 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 14 papers.
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14 citing papers in PubMed, 22 citations in OpenAlex.
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- Microbiome alterations are related to an imbalance of immune response and bacterial translocation in BDL-rats.Iranian journal of basic medical sciences · 2020Article
- Selection of Reliable Reference Genes for Analysis of Gene Expression in Spinal Cord during Rat Postnatal Development and after Injury.Brain sciences · 2019Article
- Repeated anodal trans-spinal direct current stimulation results in long-term reduction of spasticity in mice with spinal cord injury.The Journal of physiology · 2019Article
- Modulation of the expression of genes related to the system of amyloid-beta metabolism in the brain as a novel mechanism of ceftriaxone neuroprotective properties.BMC neuroscience · 2018Article
- Resistance exercise decreases heroin self-administration and alters gene expression in the nucleus accumbens of heroin-exposed rats.Psychopharmacology · 2018Article
- Phosphorylation of the Unique C-Terminal Tail of the Alpha Isoform of the Scaffold Protein SH2B1 Controls the Ability of SH2B1α To Enhance Nerve Growth Factor Function.Molecular and cellular biology · 2018Article
- Prolonged Periods of Social Isolation From Weaning Reduce the Anti-inflammatory Cytokine IL-10 in Blood and Brain.Frontiers in neuroscience · 2018Article
- Determination of stable reference genes for RT-qPCR expression data in mechanistic pain studies on pig dorsal root ganglia and spinal cord.Research in veterinary science · 2017Article
- Changes in the disposition of substance P in the rostral ventromedial medulla after inflammatory injury in the rat.Neuroscience · 2016Article
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Authors and funding
6 authors at 1 institution in 1 country.
Funding
Abstract
backgroundReal-time quantitative PCR (qPCR) is a technique frequently used to measure changes in mRNA expression. To ensure validity of experimental findings, it is important to normalize the qPCR data to reference genes that are stable and unaffected by the experimental treatment to correct for variability among samples. Unlike in some models of neuropathic pain, reference genes for models of inflammatory injury have not been validated. This study examined four candidate reference genes in an effort to identify and validate optimal genes for normalization of transcriptional changes occurring in the dorsal horn of the spinal cord and the rostral ventromedial medulla (RVM) following intraplantar injection of complete Freund's adjuvant (CFA).
resultsThe expression of hypoxanthine phosphoribosyltransferase 1 (Hprt1), beta-actin (Actb), mitogen-activated protein kinase 6 (Mapk6), and beta-2-microglobulin (B2m) was quantified in the dorsal horn and RVM of rats four days or two weeks after intraplantar injection of CFA or saline. The range of expression levels among these four genes differed by as much as 16-fold within the dorsal horn and the RVM. All four of these reference genes were stably expressed in both tissues and did not differ between saline and CFA-treated animals. Analyses using the statistical algorithms in geNorm and NormFinder programs determined that Mapk6 was the most stable gene and recommended the combination of Mapk6 and Actb, or Mapk6 and Hprt1, in such experimental conditions.
conclusionsThis study validated the four genes Hprt1, Actb, Mapk6 or B2m and showed that any one or combination of two of them are good reference genes for normalization of mRNA expression in qPCR experiments in the spinal cord and RVM in the CFA model of inflammatory injury.
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