成長期ラット顎関節円板におけるversican isoform mRNAの発現 : TaqMan probeを用いたreal-time quantitative PCRによる分析

概要

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Versican is a large chondroitin sulphate proteoglycan distributed in a variety of interstitial connective tissue. Versican is thought to be involved in extracellular matrix assembly and various biological cellular interactions. This study examined mRNA expression of versican isoforms in the temporomandibular joint (TMJ) disc of growing rats using a real-time quantitative PCR (QPCR). Male Wistar rats of 2, 4, 8, 16 and 24 weeks of age were used. Total RNA was isolated from the TMJ discs, and cDNA was generated by reverse transcriptase (RT) reaction. The quantitative analysis of mRNA expression of the target genes, QPCR, was performed using sequence specific TaqMan probes. The copy number of versican was calculated by an external standard (plasmid including target cDNA diluted serially in log steps from 10^8 to 10 copies) and was normalized to a reference cDNA, GAPDH. The QPCR showed that expression for versican mRNA was highest at 2 weeks and decreased thereafter. With respect to mRNA expressions for each versican isoform, the mRNA expression of VO, V1, and V3 gradually decreased with the growth, while the expression of V2 increased up to 8 weeks and decreased after that. The mRNA expression of each versican isoform showed age-related changes in the TMJ discs of growing rats.
北海道医療大学の論文
2005-06-30