Identification of Differentially Expressed mRNAs of the Autologous Vein Graft After an Arterial Bypass in Mongrel Dogs

Abstract

Purpose. Intimal hyperplasia (IH) is characterized by vascular smooth muscle cell (VSMC) proliferation in the intima and subsequent accumulation of extracellular matrix. A variety of factors that might be considered as possible VSMC mitogens induce the specific gene expression of VSMC. This study was designed to identify differentially expressed mRNA by method using an mRNA differential display.

Methods. A bilateral femoropopliteal reverse saphenous vein bypass was performed on both hind limbs of mongrel dogs. At 4 and 8 weeks after the implantation, the vein bypass grafts were harvested. The total RNAs were extracted from each specimen and transcribed into the cDNAs. Amplified cDNAs using 16 sets of primers were separated on DNA sequencing gel. Differentially displayed bands were excised from the gel, cloned, and then sequenced. The sequences were compared with the National Center of Biotechnology Information nonredundant sequence database using the Basic Local Alignment Search Tool (BLAST) program.

Results. Forty-three bands were cloned and sequenced. A computer search revealed that 11 of them had similarities to known genes while 32 cDNAs had no similarities to any registered DNA sequence.

Conclusions. We found several genes to be related to IH. Some of them had already had their function and sequence identified whereas some of them have yet to be identified. Further studies are necessary to determine the precise relationship between these genes and IH.