Forsøksdyr: Generation and characterization of EndoV and ApoE double KO mice (2019)

Godkjenningsdato 03.01.2020

Godkjenningsperiode 03.01.2020-02.01.2024

1. Endonuclease V (ENDOV) orthologs are conserved among all sequenced eukaryotes from yeast to humans. ENDOV is transcribed in various tissues and the protein localizes to the cytoplasm and nucleoli in human cells. ENDOV cleaves different RNA substrates with inosin (in vitro), but the molecular function of and the biological significance ENDOV in mammalian cells is not known. Initial studies show that ENDOV is highly expressed in human atherosclerotic plaques, and we hypothesize that ENDOV is important in the development of this disease. Furthermore, EndoV KO mice develop fatty liver and have increased blood glucose, a phenotype which correlates with metabolic syndrome, and increases the risk of cardiovascular disorders.
2. In this present study, we will study the metabolism and atherosclerosis development in mice lacking EndoV. The mice will be exposed for diet-induced atherosclerosis and obesity, and we will perform glucose- and insulin tolerance tests and collect blood samples for metabolite analysis. Only small volumes of blood will be collected through pinprick of tail- or calf vein, thus we estimate the stress to be mild.
3. A correlation between altered A-to-I mRNA editing and development of cardiometabolic disorders is an undescribed mechanism. This novel knowledge will open potential new therapeutic strategies.
4. For colony maintenance and generation of adequate number of experimental mice, we estimate 410 WT (C57BL/6), 410 EndoV KO, 320 ApoE KO and 320 EndoV/ApoE double KO mice – 1460 in total.
5. The interplay between individual cells, tissues and organs, and their obvious impact on the whole organism and its phenotype cannot be properly studied outside a living animal. Considerable individual variations are to be expected for cardio-metabolic studies in mice. The number of mice chosen for this project will allow us freedom to terminate animals showing discomfort, sickness or injuries without compromising the statistical validity of our study. The mice will be checked on a daily basis by KPM staff, and monitored on a weekly basis by project researchers. We will be using the form "pain assessment" (see attachment) when we monitor the animals. The mice will be housed in standard cages with nesting material and a place to hide. The animals will be kept in social groups. The experiments regarding the fatty liver phenotype and atherosclerosis will be performed after the animals have been sacrificed.