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. 2016 Sep 28;36(39):10097–10115. doi: 10.1523/JNEUROSCI.0635-16.2016

Figure 6.

Figure 6.

The SK portion of the oeAHP requires SERCA-dependent Ca2+ stores and CICR but not IP3. A, An example of IoeAHP recorded with okadaic acid (25 nm) in the pipette solution. AHP currents were evoked with a 5 pulse protocol before and after orexin-A (300 nm). B, Current-clamp recordings of the AHP evoked by the 5 spike protocol before (black), following SERCA-dependent Ca2+ store depletion with CPA (red) and during Orx A (300 nm) application in CPA (blue). C, Summary of the effect of CPA treatment on orexin-evoked changes in amplitude (Amp) and duration (50% recov) of the AHP. Data (mean ± SEM) are reported as percentage of the AHP measured in CPA before orexin application (control; dotted line = 100%). D, Summary of voltage-clamp experiments following CPA treatment. IoeAHP amplitude (mean ± SEM) was measured in CPA and CPA following blockage of SK currents with apamin (CPA w Apa). Dotted lines indicate mean ± SEM of the IoeAHP recorded under control conditions. *p < 0.05, significant difference from control. E, Examples of IoeAHP recorded with IP3R inhibitors XeC (1 μm) and 2-APB (50 μm) and the ryanodine receptor antagonist RuR (100 μm) included in the pipette solution. Calibration bars for left trace apply to the other two traces. F, Summary of IoeAHP amplitude (mean ± SEM) for recordings made with XeC, 2-APB, RuR, and RuR with apamin pretreatment. *p < 0.05, significant difference compared with Control (mean ± SEM, dotted lines) by post hoc tests following an ANOVA. G, Example membrane currents from a DR neuron (holding potential = −65 mV). The neuron was stimulated by IP3 uncaging with full-field UV flashes (500 ms; left traces) or by the 5 pulse protocol (right traces) both before (red) and after (blue) orexin-A (300 nm). Arrowhead indicates 0 pA. Calibration applies to both pairs of traces. H, Example whole-cell recording from a hippocampal CA1 neuron. Identical IP3 uncaging (UV) evoked an outward current. Calibration in G applies to H. I, Summary of evoked currents by UV uncaging of IP3 in DR neurons before (Control) and after orexin-A application (Orx A) and in hippocampal CA1 neurons (Hipp). *p < 0.05.