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Expression of Cell-Cycle-Regulating Genes in the Development of Atherosclerosis in Japanese Quail (Coturnix japonica)

M. Inafuku^*, T. Toda^,1, T. Okabe, A. Shinjo, H. Iwasaki and H. Oku

^* United Graduate School of Agricultural Sciences, Kagoshima University, Korimoto, 890-0065, Japan; and Department of Clinical Laboratory Medicine, School of Medicine, Laboratory of Plant and Animal Bleeding, Department of Bioproduction, Faculty of Agriculture, and Division of Molecular Biotechnology, Center of Molecular Bioscience, University of the Ryukyus, Nishihara, Okinawa 903-0213, Japan

¹ Corresponding author: b985703{at}med.u-ryukyu.ac.jp

The levels of mRNA expression in regulatory genes that are involved in the pathological changes of aortic atherosclerotic and fibroblastic intimal thickening was investigated in Japanese quail. The quail were divided into a control diet group and an atherogenic diet group. The quail were euthanized at 2, 4, 8, and 12 wk after consuming either a control diet or an atherogenic diet. Thereafter, both histological and immunohistochemical studies and mRNA expression analysis of the cell-cycle-regulating genes in aortic atherosclerotic lesions were performed on selected ascending aortas and their large branches. In the atherogenic diet group, aortic lipid-containing intimal and atheromatous lesions were seen mainly at 8 and 12 wk, respectively. Semiquantitative reverse-transcription PCR was used to analyze the alterations of mRNA expression on the development of atherosclerotic lesions. Messenger RNA expression of the c-fos and c-src genes showed peak levels at 8 wk in the atherogenic diet group. However, no significant alteration of c-jun mRNA expression was noted during the entire experimental period. According to the progression of aortic atherosclerotic lesions, c-myc mRNA expression in the atherogenic diet group increased chronologically, and the highest level was observed at 12 wk. Alterations in mRNA expression of proliferating cell nuclear antigen and the p27 gene were similar to that of c-myc. The levels of c-myc, proliferating cell nuclear antigen, and p27 mRNA expression was significantly correlated with the degree of aortic atherosclerotic lesion development at 12 wk in our experiment.

Key Words: cell-cycle-regulating gene • molecular genetics • atherosclerosis • quail