Fig. 5: Endogenous enkephalin in the LH is necessary for PSS-induced maladaptive responses. | Nature Communications

Fig. 5: Endogenous enkephalin in the LH is necessary for PSS-induced maladaptive responses.

From: Lateral hypothalamic proenkephalin neurons drive threat-induced overeating associated with a negative emotional state

Fig. 5

a, b Schematic for the injection of AAV-DIO-Penk shRNA into the LH (a). Red circles denote the dissected regions used in RT-qPCR (b; n = 6, 5 mice per group). In (b), two-tailed unpaired t test, t9 = 5.589, ***p = 0.000339. c–e 2.5 h food consumption test (n = 6, 7, 6 mice per group). In (d), one-way ANOVA (F(2,16) = 16.713, p < 0.001) was followed by Fisher LSD post hoc test for multiple comparisons; ***p < 0.001, †††p < 0.001 compared with eGFP-expressing PSS mice. f Representative images showing c-fos immunoreactivity and eGFP in the LH of Lepr-Cre and Penk-Cre mice. Arrowheads indicate the colocalization of c-fos with eGFP. Scale bars, 20 μm. g, h Quantification of the proportion of c-fos+ cells among eGFP-labeled LH neurons (g) or c-fos+ cells in the LH (h) (n = 6, 6, 5 mice for each PSS-NC group; n = 5, 7, 7 mice for each PSS-HFD group). In (g), two-way ANOVA (F(2,30) = 8.543, p = 0.001) was followed by Fisher LSD post hoc test for multiple comparisons; ***p < 0.001, †††p < 0.001 compared with Penk-Cre: DIO-eGFP at PSS-HFD. In (h), two-way ANOVA (F(2,30) = 11.261, p < 0.001) was followed by Fisher LSD post hoc test for multiple comparisons; *p = 0.034, ***p < 0.001 compared with mice at respective PSS-HFD conditions; †††p < 0.001 compared with Penk-Cre: DIO-Penk shRNA at PSS-HFD. i, j Schematic for CRISPR virus injections into the LH (i). Red circles denote the dissected LH used in RT-qPCR (j; n = 5, 4 mice per group). In (j), two-tailed unpaired t test, t7 = 3.631, **p = 0.00839. k, l 2.5 h food consumption test (n = 8, 6, 6 mice per group). In (k), one-way ANOVA (F(2,17) = 15.477, p < 0.001) was followed by Fisher LSD post hoc test for multiple comparisons; ***p < 0.001, †††p < 0.001 compared with PSS: Cre-. m, n Time spent (%) in PSS-paired side of mice receiving CRISPR virus in the LH (n = 6 mice). Box–whisker plots display median (center) and 2.5 to 97.5 percentiles of the distribution (bounds) with whiskers extending from min to max values. In (m), two-tailed paired t test, t5 = 3.892, *p = 0.0115; In (n), two-tailed paired t test, t5 = 0.186, p = 0.86. n.s not significant. Data are expressed as mean ± SEM.

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