Gpr126 is essential for placental development

Gpr126 is an adhesion G protein-coupled receptor (GPCR) that is expressed in the endocardium of the heart and its inactivation in mice leads to embryonic lethality and defective ventricular trabeculation. However, the mechanistic bases of this phenotype are unknown. We generated a series of standard, conditional loss- and gain-of-function alleles in mice and zebrafish, and found that cardiac abnormalities associated with global Gpr126 inactivation are secondary defects that arise as a result of placental insufficiency. Overall design: Total RNA was extracted from E12.5 mouse embryonic hearts after removing atria and the valve region in sterile ice-cold PBS. 4 pools of Gpr126+/+ (WT) and 4 pools of Gpr126?7/?7 (KO) embryonic ventricles (4 ventricles per pool) were used. The tissue was homogenized in TRI ReagentTM Solution (Invitrogen, AM9738) using steel beads in the TissueLyser LT Adapter for 5 minutes at 50 Hz. For RNA extraction, the PicoPureTM RNA Isolation kit (Applied Biosystems, KIT0204) was used according to the manufacturer's instructions. RNA was quantified and its purity checked with a NanoDrop ND-1000 spectrophotometer (Thermo Scientific). RNA integrity was verified with an Agilent 2100 Bioanalyzer instrument (Agilent Technologies).