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. 2023 Mar 17;14:1486. doi: 10.1038/s41467-023-37044-4

Fig. 6. NPY increases LepR neuron activity via disinhibition of GABAergic interneuron in the LH.

Fig. 6

a, d Representative image of GCaMP6s-expressing LHLepR neurons during the application of NPY (a) or NPY + Antagonist (d) using brain slice calcium imaging. The degree of colour brightness represents the degree of cell activity (max Z-score). Scale bar: 50 μm. The experiment was repeated 7 times (a) or 9 times (d) independently with similar results. b, e Representative traces of calcium activity of LHLepR neurons marked in (a or d). Calcium activity aligned to application of NPY (b) or NPY + Antagonist (e). Dotted black line is start of NPY application. Application scheme is shown on top. c, f Heatmap depicting calcium signals aligned to application of NPY (c) or NPY + Antagonist (f). g Average Z-score from the LHLepR calcium signal aligned to application of NPY (red) and NPY + Antagonist (purple). h, i Quantification of AUC (h) and max Z-score (i) before and after application of NPY or NPY + Antagonist. 68 cells from 7 slices (a), 70 cells from 9 slices (d). Two-sided paired t-test; ***p = 0.0002 (h, left), *p = 0.019 (i, left), two-sided unpaired t-test; *p = 0.014 (h, right), **p = 0.002 (i, right). j Quantification of the percentage of active cells. Two-sided unpaired t-test; *p = 0.014. k Representative images of td-Tomato-expressing LHLepR neurons during brain slice whole-cell recording. 3 V, the 3rd ventricle. l Representative traces of spontaneous inhibitory postsynaptic current comparing ACSF (top) and NPY (bottom). m–p Time course of sIPSCs frequency (m), amplitude (n) and quantification of normalised sIPSCs frequency (o), amplitude (p) in the last 1 min after the ACSF or NPY application. td-Tomato-expressing LHLepR neurons; 8 cells from ACSF, 6 cells from NPY. Two-way repeated measures ANOVA followed by Bonferroni post hoc test (m, n); ***p = 0.0005 (m), two-sided paired t-test (o, p); *p = 0.043 (o, right). Data are mean ± s.e.m. See Supplementary Table 1 for statistics. Source data are provided as a Source data file.