Fig. 6: GPR139-mediated dynorphin signaling in native neurons and intact circuits.
From: Homeostatic scaling of dynorphin signaling by a non-canonical opioid receptor
a Experimental design of recording calcium imaging utilizing jGCaMP7s sensor in medial habenula (MHb) region of mice. b Average of calcium response in MHb neurons of WT and Gpr139 KO mice after puff application with 100 μL of 160 μM Dyn A17 (WT n = 6 cells from 4 mice; WT/JNJ-3792165 n = 7 cells from 4 mice; Gpr139 KO, n = 6 cells from 4 mice). c Quantification of area under curve of calcium response in (b). n = 6 cells from 4 mice for WT; n = 7 from 4 mice for WT/JNJ-3792165, and n = 6 cells from 4 mice for Gpr139 KO. d Representative traces (n = 8 cells from WT mice and n = 11 cells from Gpr139 KO mice) of spontaneous firing activity of MHb neurons in brain slices from WT and Gpr139 KO mice with and without 500 nM Dyn A application. e Time course of normalized firing frequencies of MHb neurons at baseline and following Dyn A application. f Quantification of maximal drug effect as the averaged normalized firing frequency during min 9-10 (WT n = 8 cells from 4 mice; Gpr139 KO n = 11 cells from 5 mice). g Schematic of viral targeting of jGCaMP7s and Flex-ChrimsonR-tdTomato to the nucleus accumbens (NAc) and optogenetic stimulation. ISI inter-stimulation interval. h Average jGCaMP7s responses to optical stimulation in the presence or absence of 10 μM JNJ-3792165 in Pdyn+ that express ChrimsonR-tdTomato (black, n = 4 cells from 3 mice; maroon, n = 3 cells from 3 mice) and in Pdyn- neurons that do not express ChrimsonR-tdTomato (n = 7 cells from 4 mice). i Quantification of the area under the curve calculated from traces in (h). n = 4 cells from 3 mice for Pdyn+ Block; n = 3 cells from 3 mice for Pdyn+ Control. Data are mean ± SEM. Data were analyzed by (c) One-way ANOVA with Dunnett’s post hoc test, (e) Two-way ANOVA with Holm-Šídák’s test, (f) unpaired two-tailed t-test or (i) paired two-tailed t-test. *p < 0.05, **p < 0.01.
