Mechanical stimulation induces preprotachykinin gene expression in osteoarthritic chondrocytes which is correlated with modulation of the transcription factor neuron restrictive silence factor

Neuropeptides. 2008 Oct-Dec;42(5-6):681-6. doi: 10.1016/j.npep.2008.09.004. Epub 2008 Nov 5.

Abstract

We have previously demonstrated that the transcription factor termed neuron restrictive silencer factor (NRSF) and the truncated splice variant, NRSF short form (sNRSF) are major modulators of preprotachykinin A (TAC1) gene expression. In this communication we addressed whether TAC1 gene expression would be effected in response to mechanical stimulation of both normal and osteoarthritic (OA) chondrocytes. Chondrocytes were mechanically stimulated for 20 min, and then incubated under normal tissue culture conditions for 1 or 3h. RT-PCR and quantitative PCR (qPCR) were used to investigate expression of TAC1, NRSF and sNRSF mRNA at these time points. Western blotting was used to validate and confirm expression of sNRSF protein in chondrocytes in response to mechanical stimulation. We observed that TAC1 was expressed in normal chondrocytes, with no evidence of NRSF or sNRSF expression. TAC1 mRNA expression did not significantly change following mechanical stimulation in normal cells. OA chondrocytes expressed TAC1 and sNRSF mRNA, though not NRSF, and following mechanical stimulation there was a significant upregulation of both TAC1 and sNRSF mRNA, which returned to baseline levels 3h post-stimulation. sNRSF protein was upregulated at 1 and 2h following stimulation of OA chondrocytes. In summary, differential expression of TAC1 and sNRSF in OA chondrocytes associates their expression with the disease. The change in expression of sNRSF and TAC1 mRNA following mechanical stimulation in OA but not normal chondrocytes suggests that sNRSF may be involved in the regulation of SP production in OA cartilage. These differences between normal and OA mechanotransduction responses may be important in the production of phenotypic changes present in diseased cartilage.

MeSH terms

  • Aged
  • Aged, 80 and over
  • Cells, Cultured
  • Chondrocytes / cytology
  • Chondrocytes / physiology*
  • Gene Expression Regulation*
  • Humans
  • Mechanotransduction, Cellular / physiology*
  • Osteoarthritis, Knee / genetics*
  • Osteoarthritis, Knee / pathology
  • Protein Precursors / genetics*
  • Protein Precursors / metabolism
  • Repressor Proteins* / genetics
  • Repressor Proteins* / metabolism
  • Stress, Mechanical
  • Tachykinins / genetics*
  • Tachykinins / metabolism

Substances

  • Protein Precursors
  • RE1-silencing transcription factor
  • Repressor Proteins
  • Tachykinins
  • preprotachykinin