Renal Glut2 knockout mice exhibit increased glucose production. In vivo increase in total (a), hepatic (b), and renal (c) glucose production through gluconeogenesis with pyruvate as a substrate in 28 weeks old male renal Glut2 knockout mice. Decreased hepatic glucose 6-phosphate (d), glucose 1-phosphate (e), and fructose 6-phosphate (f) in renal Glut2 knockout mice 12 weeks after inducing the Glut2 deficiency. *p<0.05, ***p<0.001, unpaired 2-tailed Student’s t-test. Data are presented as mean ±sem.

Enhanced activity of hypothalamic-pituitary-adrenal axis in renal Glut2 knockout mice. Representative images from fluorescence RNA in situ hybridization showing an increase in expression of corticotropin releasing hormone (Crh) in the paraventricular nucleus of the hypothalamus (which is identified here using a white oval shape) in 28 weeks old male renal Glut2 knockout mice (a). Scale, 100 µm. For the quantification shown next to the images, 4 sections per mouse and 3 areas of interest per section were analyzed in 4 mice. qRT-PCR analysis showing an increase in hypothalamic Crh (b), data from ELISA demonstrating an increase in plasma adrenocorticotropic hormone (ACTH) (c) and corticosterone (d) in 12 weeks old male renal Glut2 knockout mice 12 weeks after inducing the Glut2 deficiency. *p<0.05, **p<0.01, ***p<0.001, unpaired 2-tailed Student’s t-test. Data are presented as mean ±sem.

Effects of afferent renal denervation on blood glucose levels and hypothalamic-pituitary-adrenal axis in renal Glut2 knockout female mice. Afferent renal denervation decreases fed (random) and fasting (overnight, 6:00 pm – 9:00 am) blood glucose levels (a,b), restores expression of hypothalamic corticotropin releasing hormone (Crh) (c) measured using RT-qPCR, and plasma corticosterone (d) without affecting plasma insulin levels (e) in 30 weeks old female renal Glut2 knockout mice 16 weeks after inducing the Glut2 deficiency. **p<0.01, ***p<0.001, two-way ANOVA followed by a Tukey’s post hoc multiple comparison test. Data are presented as mean ±sem.

Changes in levels of plasma proteins in renal Glut2 knockout male mice 12 weeks after inducing the Glut2 deficiency. Representative image of two-dimensional difference gel electrophoresis with numbered protein spots of interest is shown in (a). Internal standard was prepared using equal amounts of protein of each plasma sample as a quality control (Cy2 labeled, pseudo blue), plasma proteins from control group were labeled using Cy3 dye (shown in pseudo green), and plasma proteins from renal Glut2 knockout mice were labeled using Cy5 dye (shown in pseudo red). The identified proteins and their fold change in 28 weeks old male renal Glut2 knockout mice compared to the control group is shown in (b). The number on each bar graph in (b) represents the corresponding protein spot on the gel shown in (a). MBL1, mannose binding lectin 1; Gpx3, glutathione peroxidase 3; MUP18, major urinary protein 18.