Development of CIDEA reporter mouse model and its application for screening thermogenic drugs

Abstract

Cell death-inducing DNA fragmentation factor-like effector A (CIDEA) is a lipid droplet-associated protein and is a known marker of the thermogenic capacity of brown/beige adipocytes. To monitor the expression of CIDEA in live mice in a non-invasive manner, we generated CIDEA reporter mice expressing multicistronic mRNAs encoding CIDEA, luciferase 2, and tdTomato proteins under the control of the Cidea promoter. The expression level of endogenous CIDEA protein in adipose tissue was not affected by the expression of polycistronic reporters. The two CIDEA reporters, luciferase 2 and tdTomato, correctly reflected CIDEA protein levels. Importantly, luciferase activity was induced by cold exposure and the treatment with β3-adrenergic receptor agonist CL316,243 in interscapular and inguinal adipose tissue, which was detectable by in vivo bioluminescence imaging. We further evaluated the effects of candidate brown adipogenic agents using this CIDEA reporter system and demonstrated a positive correlation between drug-induced luciferase activity and thermogenic gene expression levels both in vitro and in vivo. Collectively, we established a dual CIDEA reporter mouse model in which fluorescence and luminescence signals correctly reflect CIDEA expression, and therefore, suggested that this reporter system can be used to evaluate the thermogenic efficacy of candidate molecules.

Anti-obesity effects of heat-transformed green tea extract through the activation of adipose tissue thermogenesis

“…CIDEA reporter mice were used to determine the most effective treatment dosage. CIDEA reporter mice were previously developed to monitor CIDEA expression through fluorescence and luminescence signals [ 19 ]. The effects of high-dose (HTGT + EMIQ: 100 mg/kg each) and low-dose (HTGT + EMIQ: 50 mg/kg each) administration were compared, and mirabegron (10 mg/kg), a well-known beta-3 adrenergic receptor (β3-AR) agonist, was used as a positive control.…”

“…6-week-old male C57BL/6 mice (JA Bio, South Korea) and Cell death-inducing DNA fragmentation factor-like effector A (CIDEA) reporter mice (Cidea tm1(Luc2/tdTomato)Yjl /KMPC) were used for in vivo experiments following approved protocols by Institutional Animal Care and Use Committees of Seoul National University (SNU-200904-7-3, SNU-201221-3). CIDEA reporter mice were obtained as described previously [ 19 ]. All experiments were performed at Animal Center for Pharmaceutical Research in Seoul National University, where mice were housed at 22 ± 1 °C, 12-h light/12-h dark cycle condition with free access to food and water.…”

“…In vivo bioluminescence imaging was performed as described previously [ 19 ]. Briefly, the region of interest of CIDEA reporter mice was shaved with an electric razor.…”

Anti-obesity effects of the dual-active adenosine A2A/A3 receptor-ligand LJ-4378

“…Animal experiments were performed following approved protocols by Institutional Animal Care and Use Committees of Seoul National University (SNU-201107-1, SNU-201221-3). C57BL/6 male mice and CIDEA reporter mice [ 17 ] were used (8-week-old, male). Mice were housed at 12 h-light/12 h-dark cycle condition with free access to a normal chow diet (NCD, Purina Lab, 38057, protein: 24.52% calories, carbohydrates: 63.07% calories, fat: 12.41% calories, Seongnam, Republic of Korea) and water at 22 ± 1 °C.…”

“…Western blot analysis and qPCR were conducted, as described previously [ 17 ]. Primers used for the qPCR analyses are listed in Supplementary Table S1 .…”