Figure 4: Persistent shifts in EGABA induced by Cl-out4 activation in cells with compromised cation-chloride co-transport.
From: Cl-out is a novel cooperative optogenetic tool for extruding chloride from neurons
(a) Example recordings showing repeated muscimol applications, either side of a period of intense Cl-out activation, in cells bathed in VU0463271 and bumetanide to block KCC2 and NKCC1 respectively. A test muscimol application was delivered every 20 s, because the response was stable at that frequency in baseline conditions. Note the progressive shift in the voltage response during the reiterative light activation, reflecting the shifting ChloC/GtACR component of the current as ECl shifts more negative. (b) Pooled data of the IPSP shift, normalized to the maximal shift, immediately after the period of opsin activation (mean± s.e.m.). (c) Pooled data of the time constant for recovery of GABA−ΔV to baseline levels, showing a significant additional slowing of recovery when both KCC2 and NKCC1 were blocked (τ(VU+Bume)=58.2±11.5 s, n=7), over the situation where only KCC2 was blocked (τ(VU)=37.5±6.7 s, n=7; τ(VU (Bume washout)=33.1±5.1 s, n=5).
