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. 2014 Dec 23;8:145. doi: 10.3389/fncir.2014.00145

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Copyright © 2014 Nerlich, Keine, Rübsamen, Burger and Milenkovic.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution and reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Asynchronous release influences IPSC decay in SBCs. (A) Superimposed responses to a 100 Hz stimulation in 2 mM calcium- and 8 mM strontium-ACSF. Right: Normalized last IPSCs showing an increased incidence of asynchronous release events under strontium. (B1) Summary histograms of asynchronous release events, before and after a 10 pulse stimulation at 100 Hz for both conditions (n = 6). (B2) Mono-exponential fits to the data in B1. The amount of asynchronous events after stimulation was increased and the time of increased incidence prolonged under 8 mM Sr²⁺ (A and τ: Ca²⁺ vs. Sr²⁺ p < 0.05). (C) Substitution of Ca²⁺ by Sr²⁺ prolonged the weighted decay time of IPSCs after 100 Hz and 333 Hz stimulation, but not of single IPSCs (n = 6, single: p = 0.8, 100 Hz: p < 0.05, 333 Hz: p < 0.01, RM ANOVA). (D) Averaged trains of 50 IPSCs at 333 Hz before and after bath application of 100 μM EGTA-AM. Right: normalized last IPSCs. (E) Summary histograms of asynchronous release events before and after application of EGTA-AM. Note the lack of the increase in asynchronous release after stimulation in EGTA-AM (n = 5, bin width: 20 ms; control: z = 29.2 p < 0.001; +EGTA-AM: z = −1.01 p = 0.16, z-test). (F) EGTA-AM shortened τ_wd by about 30 % regardless of stimulation frequency (n = 5, *p < 0.05, ***p < 0.001, RM ANOVA). (G) EGTA-AM equally affects the control (mixed glycine/GABA) IPSCs, isolated glycinergic (+20 μM SR 95531), and GABAergic IPSCs (+0.5 μM strychnine) in terms of decreasing the weighted decay time constants (co-transmission: n = 5, p < 0.001; glycinergic: n = 5, p < 0.05; GABAergic: n = 5, p < 0.01; ANOVA).