SRA

At 2 months of age, liver-specific insulin receptor knockout (LIRKO) mice present hyperglycemia and hyperinsulinemia. Furthermore, LIRKO mice have increased levels of hepatic cholesterol. Indeed, many changes seen in cholesterol metabolism in LIRKO mice are also observed in humans with metabolic syndrome. For example, both show decreased levels of HDL and increased secretion of apoB and VLDL. These findings make the LIRKO mouse a unique non-dietary model of insulin resistant, hyperglycemia, dyslipidemia and atherosclerosis that resembles several clinical features of the human metabolic syndrome. By hepatic transcriptomic analysis of the wild-type (WT) offspring of LIRKO mice, we identify that members of the TGF-ß family are differentially expressed in the offspring, including the NREP and GDF15. Overall design: LIRKO (insulin receptor-IRlox/lox; Albumin-Cre+/-) mice were generated as previously described. The control offspring group consisted of the F1 offspring from a control male and female (insulin receptor-IRlox/lox; Albumin-Cre-/-). Control parents were crossed for 4 generations for minimizing any epigenetic memory from the presence of Cre. Father LIRKO offspring (FL) resulted from the crossing of a male LIRKO with a control female. Mother LIRKO offspring (ML) resulted from the breeding of a control male with a LIRKO female. Liver total RNA was extracted using standard Trizol reagent (Invitrogen) and cleaned up using RNeasy mini kit columns (Qiagen). Polyadenylated RNAs were isolated using NEBNext Magnetic Oligo d(T)25 Beads and first-strand cDNA synthesis was performed using NEBNext RNA first-strand synthesis module (New England BioLabs). Directional second strand synthesis was performed using NEBNExt Ultra Directional second strand synthesis kit. The NEBNext DNA Library Prep Master Mix Set for Illumina was then used to prepare individually bar-coded next-generation sequencing expression libraries. Paired-end sequencing was performed on an Illumina HiSeq2500 sequencer.