The identification of glucose-responsive genes in the skeletal muscle of WT and ob/ob mice.
The ability to produce stable blood glucose is indispensable for human life and health. The mechanisms regulating glucose-responsive metabolism in the skeletal muscle and how these mechanisms are altered in obesity have yet to be identified. Here we measured transcriptomic data in the skeletal muscle of wild type (WT) and leptin-deficient obese (ob/ob) mice following oral glucose administration, and identified the genes that were quantitatively changed by oral glucose administration. After overnight fasting (16 h), ten-week-old male C57BL/6 wild-type and ob/ob mice were administered 2 g/kg body weight of glucose orally. At 0, 20, 60, 120, 240 min after administration, mice were sacrificed by cervical dislocation and the skeletal muscle was dissected and immediately frozen in liquid nitrogen. The frozen skeletal muscle was pulverized with dry ice to a fine powder with a blender and separated into tubes. Total RNA was extracted from the skeletal muscle using RNeasy Mini Kit (QIAGEN) and QIAshredder (QIAGEN) and assessed for quantity using Nanodrop (Thermo Fisher Scientific) and for quality using the 2100 Bioanalyzer (Agilent Technologies). cDNA libraries were prepared using SureSelect strand-specific RNA library preparation kit (Agilent Technologies). The resulting cDNAs were subjected to 100-bp paired-end sequencing on the Illumina HiSeq2500 Platform (Illumina).