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. 2018 Feb 7;38(6):1588–1599. doi: 10.1523/JNEUROSCI.1925-17.2017

Figure 5.

Figure 5.

Effects of dynorphin on the LH/PFVgat → Orx input. A, DynA (500 nm) reduces the amplitude of the photo-evoked IPSCs, and this effect does not desensitize during 10 min DynA application. Con, 30 photo-evoked IPSCs recorded just before DynA application and average photo-evoked IPSC; DynA (5 min), 6 photo-evoked IPSCs recorded between 4 and 5 min of a 10 min DynA application and average photo-evoked IPSC; DynA (10 min), 6 photo-evoked IPSCs recorded between 9 and 10 min of a 10 min DynA application and average photo-evoked IPSC; wash, 30 photo-evoked IPSCs recorded after 10 min washout and average photo-evoked IPSCs. Gray represents individual photo-evoked IPSCs. Black represents average photo-evoked IPSCs. B, The effect of DynA is abolished by the κ-opioid receptor antagonist nor-BNI (1 μm). Gray represents 30 photo-evoked IPSCs. Black represents average photo-evoked IPSCs. C, Summary graph representing the DynA-mediated inhibition in control ACSF after 5 min application of DynA (5 min; n = 7), after 10 min application of Dyn-A (10 min; n = 7), and in the presence of nor-BNI (nor-BNI + DynA; n = 5, one-way ANOVA, F = 11.00, p = 0.001; Fisher's LSD post hoc test, **p < 0.01, comparing both DynA 5 min and DynA 10 min vs nor-BNI + DynA). D, E, DynA has no significant effect on the photo-evoked IPSC latency (D, n = 7; one-way ANOVA, F = 0.146, p = 0.866), or photo-evoked probability (E, n = 7; one one-way ANOVA, F = 1.084, p = 0.359). F, G, In the same orexin neurons in which DynA inhibits the LH/PFVgat → Orx input, DynA also reduces the sIPSC frequency. Cumulative distribution plots of sIPSC interevent intervals compiled from 7 orexin neurons (F; 200 ms bins; two-way ANOVA F = 4.606, p < 0.001; Bonferroni's multiple-comparisons post hoc test, **p < 0.01, comparing control and DynA). Mean values of the sIPSC frequency (G; n = 7; one-way ANOVA, F = 4.956, p = 0.019; **p < 0.01, Fisher's LSD post hoc test). Photo-evoked IPSCs were evoked by 10 ms light pulses and were recorded at Vh = 0 mV using the Cs-methane-sulfonate-based pipette solution.