RNA-seq Analysis of Gs-linked GPCRs expressed in mouse inguinal white adipocytes (iWAT), epididymal white adipocytes (eWAT) and brown adipose tissues (BAT)

Purpose: The goals of this study are to identify Gs-linked GPCRs that are endogenously expressed by mouse adipose tissue, we subjected RNA prepared from isolated mouse adipocytes (iWAT and eWAT) and BAT tissue to RNA-seq analysis. Methods: Total RNA extracted from mature adipocytes (iWAT and eWAT) and BAT tissue of C57BL/6J mice (16-week old males) maintained on regular chow were used to construct high throughput sequencing libraries. RNAs with RIN >8 (assessed by the Agilent 2100 Bioanalyzer system) were used to prepare transcriptome libraries using the NEBNext Ultra RNA library prep kit (New England Biolabs). High throughput RNA-sequencing was performed using a HiSeq 2500 instrument (Illumina) at the NIDDK Genomic Core Facility (NIH, Bethesda, MD). Raw reads were mapped to the mouse (mm9) genome. GPCRs were extracted from the RNA-seq data using R form. Gs-coupled GPCRs were identified using the IUPHAR/BPS Guide to Pharmacology Database (https://www.guidetopharmacology.org/). Results: Our study demonstrated that mouse adipocytes/BAT tissue express several GPCRs that are selectively coupled to Gs, including the V2 vasopressin receptor, the glucagon receptor, and different melanocortin receptor subtypes. Inguinal white adipocytes (iWAT), epididymal white adipocytes (eWAT), and brown adipose tissues (BAT) were isolated from 16-week-old male C57BL/6J mice consuming regular chow (n=6)