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Cardiovascular disease is linked to obesity, the metabolic syndrome, and altered 24hour (circadian) rhythms. Although the underlying mechanisms remain undefined, transcriptome analysis in the heart is beginning to provide important insights into the cardiometabolic pathogenesis. The reliability and accuracy of real-time quantitative PCR generated gene expression data is largely dependent on the selection of suitable reference genes (RG), which must be constitutively expressed regardless of cardio-metabolic disease state and time of day. However, many studies do not employ the appropriate selections strategies. In this study we determined the expression stability of seven candidate RGs (GAPDH, YWHAZ, B2M, EIF4A2, ATP5?, ACTB and CYC1) in a mouse model of diet-induced metabolic syndrome in both the day and night, using geNorm qBasePLUS software. RG expression varied in hearts of normal fed versus high fat fed mice, and was also dependant on the time of day. When all experimental variables were considered YWHAZ and ACTB were ranked the most stable and therefore the most suitable genes for generating comparative gene expression data in heart tissue from murine models of cardiometabolic disease. This study provides important information for reference gene selection, and will aid further transcriptome investigations into heart organ function |
