Fig. 2
Deletion of GLP-1R expressing neurons in the DMH results in hyperphagia and obesity. A–C: An illustration of the control blank-saporin (B-SAP) or Exenatide4-saporin (Ex4-SAP) injection site (A). The sections with Nissl staining after B-SAP (B) or Ex4-SAP (C) injection. D–H Relative mRNA expression of GLP-1R (D), GAD67 (E), NPY (F), CCKA receptors (CCKAR) (G) and leptin receptors (LEPR) (H). I–N Colocalization of GLP-1R (I, L) and GAD67 (J, M) in the DMH. K and N indicate the merged image of I and J, L and M, respectively. l–N indicate enlarged image of dashed square in I–K, respectively. Scales in I–K = 50 μm. Scales in L–N = 10 μm. O Twenty-four hours of locomotor activity (count per hour) of rats injected with control B-SAP or Ex4-SAP into the bilateral DMH. (n = 9, 7) P The counts of locomotor activity (Left), and the ratio of locomotor activity (Right) between the dark phase (DP) and light phase (LP) in B-SAP- and Ex4-SAP-injected rats. * P < 0.05, ** P < 0.01. Student t-test. (n = 9, 7). Q Twenty-four hours of food intake in rats that were injected with B-SAP or Ex4-SAP. *P < 0.05, **P < 0.01. Two-way ANOVA. (n = 9, 6). R The amount of food intake (Left) and the ratio of food intake (Right) between the dark phase (DP) and light phase (LP) in the B-SAP- and Ex4-SAP-injected rats. * P < 0.05, ** P < 0.01. Student’s t-test. (n = 9, 6). S: Daily food intake for 4 weeks after B-SAP or Ex4-SAP injection into the DMH. *P < 0.05, **P < 0.01. Two-way ANOVA. (n = 6, 6) T Cumulative food intake for 4 weeks in the B-SAP- and Ex4-SAP-injected rats. *P < 0.05. Student’s t-test. (n = 6, 6). U Change of BW for 4 weeks after B-SAP or Ex4-SAP injection. ** P < 0.01. Two-way ANOVA. (n = 6, 6). V The comparison of body size of the B-SAP- and Ex4-SAP-injected rats at 28 days after injection. Left: B-SAP, Right: Ex4-SAP. W: Food intake for 2 h after refeeding in B-SAP or Ex4-SAP injected rats. (n = 9, 8). X, Y Representative c-Fos expression after refeeding in the SCN of rats in which B-SAP (X) or Ex4-SAP (Y) was injected into the DMH. Scale bars = 100 μm. Z The number of c-Fos-positive neurons in the SCN per section after being refed. * P < 0.05, Student’s t-test. (n = 9, 8)