Identification of robust cardiac reference genes in a mouse model of cardiometabolic disease

Abstract

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

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Identifiers

ORCID for Kerry Hyde: orcid.org/0000-0003-4841-5953

ORCID for Mark A. Hanson: orcid.org/0000-0002-6907-613X

ORCID for Christopher D. Byrne: orcid.org/0000-0001-6322-7753

ORCID for Felino R. Cagampang: orcid.org/0000-0003-4404-9853

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