Expression stability of commonly used reference genes in canine articular connective tissues

Fiona Marriage, Philip Day, Duncan Ayers, Dylan N. Clements, Fiona Salway, Philip J R Day

    Research output: Contribution to journalArticlepeer-review


    Background: The quantification of gene expression in tissue samples requires the use of reference genes to normalise transcript numbers between different samples. Reference gene stability may vary between different tissues, and between the same tissue in different disease states. We evaluated the stability of 9 reference genes commonly used in human gene expression studies. Realtime reverse transcription PCR and a mathematical algorithm were used to establish which reference genes were most stably expressed in normal and diseased canine articular tissues and two canine cell lines stimulated with lipolysaccaride (LPS). Results: The optimal reference genes for comparing gene expression data between normal and diseased infrapatella fat pad were RPL13A and YWHAZ (M = 0.56). The ideal reference genes for comparing normal and osteoarthritic (OA) cartilage were RPL13A and SDHA (M = 0.57). The best reference genes for comparing normal and ruptured canine cranial cruciate ligament were B2M and TBP (M = 0.59). The best reference genes for normalising gene expression data from normal and LPS stimulated cell lines were SDHA and YWHAZ (K6) or SDHA and HMBS (DH82), which had expression stability (M) values of 0.05 (K6) and 0.07 (DH82) respectively. The number of reference genes required to reduce pairwise variation (V) to
    Original languageEnglish
    Article number7
    JournalBMC Veterinary Research
    Publication statusPublished - 2007


    • Adipose Tissue
    • Algorithms
    • Animals
    • Cartilage,Articular
    • Cell Line
    • Dog Diseases
    • Dogs
    • Gene Expression
    • Gene Expression Regulation
    • genetics
    • Hindlimb
    • Human
    • Joints
    • metabolism
    • methods
    • Osteoarthritis
    • Reference Standards
    • Reverse Transcriptase Polymerase Chain Reaction
    • veterinary


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