Single-cell RNA sequencing of adipocyte-specific hM4Di-DREADD expressing mice 24 h after cardiac ischemia.
We generated mice expressing hM4Di-receptor heterozygous, tamoxifen-inducible and adipocyte-specific to inhibit adipocyte lipolysis specifically during and shortly after cardiac ischemia. Expression was induced in 10 weeks old male mice via 5d of consecutive i.p.-injections of 500µg hydroxytamoxifen/day. Cre positive, hM4Di negative male mice were used as control. After 2 weeks washout phase mice underwent closed chest ischemia with first ligature-induction surgery and 6 d later 1 hour closed chest ischemia followed by 24 h reperfusion. To inhibit lipolysis, the specific synthetic agonist of hM4Di receptor Dreadd-Agonist 21 (3mg/kg BW) was i.p.-injected before ischemia and directly after ischemia. Mice were sacrificed and hearts excised, single cells isolated to perform single cell RNA sequencing. The goal was to identify differences in gene expression due to specific peri-infarct inhibition of lipolysis in the different cell types in the heart after ischemia.