Mrpl10 and Tbp Are Suitable Reference Genes for Peripheral Nerve Crush Injury

Int J Mol Sci. 2017 Jan 27;18(2):263. doi: 10.3390/ijms18020263.

Abstract

Peripheral nerve injury triggers the dysregulation of a large number of genes at multiple sites, including neurons, peripheral nerve stump, and the target organ. Housekeeping genes were frequently used as reference genes to normalize the expression values of target genes. Suitable selection of housekeeping genes that are stably expressed after nerve injury minimizes bias elicited by reference genes and thus helps to better and more sensitively reflect gene expression changes. However, many housekeeping genes have been used as reference genes without testing the expression patterns of themselves. In the current study, we calculated the expression stability of nine commonly used housekeeping genes, such as 18S (18S ribosomal RNA), Actb (β-actin), CypA (cyclophilin A), Gapdh (glyceraldehydes-3-phosphate dehydrogenase), Hprt (hypoxanthine guanine phosphoribosyl transferase), Pgk1 (phosphoglycerate kinase 1), Tbp (TATA box binding protein), Ubc (ubiquitin C), YwhaZ (tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation), and four newly identified housekeeping genes, including Ankrd27 (Ankyrin repeat domain 27), Mrpl10 (mitochondrial ribosomal protein L10), Rictor (rapamycin-insensitive companion of mTOR, Complex 2), and Ubxn 11 (UBX domain protein 11), in both distal sciatic nerve samples and dorsal root ganglion (DRG) samples after sciatic nerve injury. Our results suggested that following peripheral nerve injury, Mrpl10 and Tbp might be used as suitable reference genes for sciatic nerve stump and DRGs, respectively.

Keywords: NormFinder; distal nerve stump; dorsal root ganglion; geNorm; housekeeping gene; rat peripheral nerve injury.

MeSH terms

  • Animals
  • Crush Injuries / genetics*
  • Female
  • Ganglia, Spinal / metabolism
  • Gene Expression Regulation
  • Genes, Essential
  • Immunohistochemistry
  • Nerve Crush*
  • Peripheral Nerve Injuries / genetics*
  • Rats, Sprague-Dawley
  • Reference Standards
  • Ribosomal Proteins / genetics*
  • Ribosomal Proteins / metabolism
  • Sciatic Nerve / injuries
  • Sciatic Nerve / metabolism
  • Sciatic Nerve / pathology
  • Software
  • TATA-Box Binding Protein / genetics*
  • TATA-Box Binding Protein / metabolism

Substances

  • Ribosomal Proteins
  • TATA-Box Binding Protein