Fig. 2: D2R and InsR expression in the CeA and regulation of InsR signaling by D2Rs. | Molecular Psychiatry

Fig. 2: D2R and InsR expression in the CeA and regulation of InsR signaling by D2Rs.

From: Dopamine D2 receptor modulation of insulin receptor signaling in the central amygdala: implications for compulsive-like eating behavior

Fig. 2

A D2R and InsR expression in the CeA of Drd2-eGFP mice. Left: Low magnification. Right: High magnification of area in the yellow box shown in the left. Red: InsR, green: Drd2-eGFP, blue: DAPI. B Quantitative analysis of D2R and InsR expression in the CeA of Drd2-eGFP mice. Drd2-eGFP n = 6 (8 sections/mouse). C InsR expression in the CeA of WT and Drd2−/− mice. Green: InsR, blue: DAPI. D Proportion of InsR-expressing cells in the CeA of WT and Drd2−/− mice. Unpaired Student’s t-test, two-tailed,***p = 0.0005. WT n = 3, Drd2−/− n = 3 (6sections/mouse). E Schematic diagram of AAV-GFP or AAV-Cre-GFP injection into the CeA of Drd2flox/flox mice. F Left: Representative images of GFP or Cre-GFP virus expression in the CeA in low magnification. Right: InsR expression in the CeA of GFP and Cre-GFP virus- injected mice as detected by immunohistochemistry. Red; InsR, Green: GFP or Cre-GFP, Blue: DAPI. Scale bar: low magnification, 100 μm, high magnification, 20 μm. G Quantitative analysis of InsR expression in the CeA. The proportion of InsR expression in GFP and Cre-GFP-injected Drd2flox/flox mice. Unpaired Student’s t-test, two-tailed, *p = 0.0123. GFP n = 3, Cre-GFP n = 3. H Expression of pInsRTyr972 with GFP or Cre-GFP-injected Drd2flox/flox mice. Saline/Insulin (10 mU)/quinpirole (1 µg) was infused 30 min before sacrifice. Left: low-magnification of the CeA of saline(S), insulin(I), or quinpirole(Q)-infused mice. Middle/Right: high-magnification of pInsRTyr972 expression in the CeA detected by immunohistochemistry. Quantitative analysis: three sections of one hemisphere/mouse, 4mice/group. Red: pInsRTyr972, Green: GFP/Cre-GFP, Blue: DAPI, Arrowhead: pInsRTyr972-positive cells. I pInsRTyr972 expression in the CeA of GFP or Cre-GFP-injected Drd2flox/flox mice. Two-way ANOVA, Bonferroni posthoc tests, group×drug interaction: F2,18 = 9.09, **p = 0.0019. GFP S/I/Q n = 4, Cre-GFP S/I/Q n = 4. J pAktSer473 expression in the CeA of GFP or Cre-GFP-injected Drd2flox/flox mice. Two- way ANOVA, Bonferroni posthoc tests, group×drug interaction: F2,26 = 5.66, **p = 0.0091, GFP S/I/Q n = 6, Cre-GFP S/Q n = 5, Cre-GFP I n = 4. K Expression of pPTP1BSer50 in CeA of GFP/Cre-GFP-injected Drd2flox/flox mice. Quantitative analysis: three sections of one hemisphere/mouse, n=4mice/group. Red: pPTP1B, Green: GFP/Cre-GFP, Blue: DAPI, Arrowhead: pPTP1B-positive cells. L pPTP1BSer50 expression. Two-way ANOVA,Bonferroni posthoc tests, group×drug interaction: F2,18 = 5.13, *p = 0.0173, GFP S/I/Q n = 4, Cre-GFP S/I/Q n = 4. Scale bars in all images: low-magnification, 100μm, high-magnification, 20μm. All values in the data represent mean ± SEM.

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