Next-Generation Sequencing Transcriptome Quantitative Analysis of Hearts of Male Fetuses From Obese Female Mice

Obesity during pregnancy increase the risk for cardiac in the offspring. We used RNA next-generation sequencing analysis to have a snapshot of the heart transcriptome in male offspring exposed to maternal obesity and weaned onto an obesogenic diet. Overall design: Female C57BL/6J mice received either commercial standard (RM1) or a high-fat diet [20% lipids (Special Dietary Services)] plus sweetened condensed milk [55% simple carbohydrates/8% lipids (Nestle, UK)] from weaning. After around 6 weeks on their respective diets, mice were mated with male counterparts and went through a first pregnancy and lactation to ensure their breeding competence. Mice were mated a second time, and first day of pregnancy was marked by the detection of a vaginal plug. On day 18.5 of gestation, pregnant mice were killed by raising CO2 concentrations or cervical dislocation. Fetuses were surgically removed and immediately killed by laying in cold buffer. Fetal heart ventricles were dissected, weighed, snap frozen and stored at -80 °C. Cardiac RNA from male offspring was extracted using a Qiazol/miRNeasy mini kit protocol (Qiagen, Hilden, Germany). Library preparation for mRNA sequencing followed manufacturer protocol of TruSeq RNA Library Preparation kit (Illumina, Cambridge, UK).