The impact of maternal diet on endocrine islet progenitor cells in mice
The goal of this study is to examine how altering maternal diet, including poor- or over-nutrition, impacts the gene expression levels in islet progenitors, including those cells that have not been committed to the endocrine fate and those having committed to it. At present, we will determine the gene expression changes in these progenitors. Overall design: Ngn3-eGFP mice was used to purifiy progenitor cells. These mice have eGFP cDNA replacing the coding regions of Ngn3. In Ngn3eGFP/eGFP embryos, the eGFP+ cells represent progenitors that are ready to become islet progenitors but have not done so. In Ngn3eGFP/+ embryos, the eGFP+ cells are committed endocrine cells. For purifying  these cells, 6-weeks old Ngn3eGFP/+ female mice was fed with control nornal dient (ND), figh-fat diet (HFD), or low protein diet (LPD) for two weeks. They were then crossed with Ngn3eGFP/+ males that have been fed with control diet. After mating, females were kept on the same diet until embryo collection at E15.5. The Ngn3eGFP/eGFP and Ngn3eGFP/+ embryos were identified via genotyping and used for panceatic tissue collection, dissociation, and FACS-sorting. eGFP+ cells were used for RNA preparation, which were subject to RNAseg using the Illumina platforms (NovaSeq 6000) in the Vanderbilt Technologies for Advanced Genomics (VANTAGE), who also provides quality control data and primary data processing for alignment and feature (gene) identification. Note that two samples were prepared in a similar fashion, except the cells were sorted from the hypothalamus of E15.5 mice.