title: An organ culture system to model early degenerative changes of the intervertebral disc II: profiling global gene expression changes.
abstract: BACKGROUND
  Despite many advances in our understanding of the molecular basis of disc degeneration, there remains a paucity of preclinical models which can be used to study the biochemical and molecular events that drive disc degeneration, and the effects of potential therapeutic interventions. The goal of this study is to characterize global gene expression changes in a disc organ culture system that mimics early nontraumatic disc degeneration.
  
  
  METHODS
  To mimic a degenerative insult, rat intervertebral discs were cultured in the presence of TNF-α, IL-1β and serum-limiting conditions. Gene expression analysis was performed using a microarray to identify differential gene expression between experimental and control groups. Differential pattern of gene expression was confirmed using quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) or Western blot.
  
  
  RESULTS
  Treatment resulted in significant changes in expression of more than 1,000 genes affecting many aspects of cell function including cellular movement, the cell cycle, cellular development, and cell death and proliferation. Many of the most highly upregulated and downregulated genes have known functions in disc degeneration and extracellular matrix hemostasis. Construction of gene networks based on known cellular pathways and expression data from our analysis demonstrated that the network associated with cell death, cell cycle regulation and DNA replication and repair was most heavily affected in this model of disc degeneration.
  
  
  CONCLUSIONS
  This rat organ culture model uses cytokine exposure to induce wide gene expression changes with the most affected genes having known reported functions in disc degeneration. We propose that this model is a valuable tool to study the etiology of disc degeneration and evaluate potential therapeutic treatments.
authors.forename: Dessislava Z
authors.initials: DZ
authors.lastname: Markova
authors.forename: Christopher K
authors.initials: CK
authors.lastname: Kepler
authors.forename: Sankar
authors.initials: S
authors.lastname: Addya
authors.forename: Hallie B
authors.initials: HB
authors.lastname: Murray
authors.forename: Alexander R
authors.initials: AR
authors.lastname: Vaccaro
authors.forename: Irving M
authors.initials: IM
authors.lastname: Shapiro
authors.forename: D Greg
authors.initials: DG
authors.lastname: Anderson
authors.forename: Todd J
authors.initials: TJ
authors.lastname: Albert
authors.forename: Makarand V
authors.initials: MV
authors.lastname: Risbud
mesh_terms: D000818:Animals; D015153:Blotting, Western; D016922:Cellular Senescence / Q000187:drug effects / Q000235:genetics; D016000:Cluster Analysis; D016895:Culture Media, Serum-Free / Q000494:pharmacology; D004195:Disease Models, Animal; D053263:Gene Regulatory Networks / Q000187:drug effects / Q000235:genetics; D006801:Humans; D053583:Interleukin-1beta / Q000494:pharmacology; D007403:Intervertebral Disc / Q000378:metabolism* / Q000473:pathology; D055959:Intervertebral Disc Degeneration / Q000235:genetics* / Q000378:metabolism; D020411:Oligonucleotide Array Sequence Analysis; D009924:Organ Culture Techniques / Q000379:methods*; D051381:Rats; D020133:Reverse Transcriptase Polymerase Chain Reaction; D059467:Transcriptome / Q000187:drug effects / Q000235:genetics*; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
chemical_list: D016895:Culture Media, Serum-Free; D053583:Interleukin-1beta; D014409:Tumor Necrosis Factor-alpha