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. 2017 Oct 11;37(41):9859–9870. doi: 10.1523/JNEUROSCI.2761-16.2017

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Copyright © 2017 Pirttimaki, Sims et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution License Creative Commons Attribution 4.0 International, which permits unrestricted use, distribution and reproduction in any medium provided that the original work is properly attributed.

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Figure 5. — SIC-induced neuronal calcium signaling. A, Fluorescence image of a cortical slice loaded with Fluo-4AM (left) and SR101. Middle, Overlaid green and red images show the area at a time at beginning of acquisition (Frame a) and during a synchronized event (Frame b), with square bounded region delineating an area exhibiting a calcium increase in cells not SR101 loaded. Right, Active regions expanded. B, Fluorescence image of a field of neurons in a cortical slice loaded with Fluo-4 AM (left) and traces of fluorescence over time with spontaneous SCRs (right) for circled neurons in the presence of TTX. Scale bar, 50 μm. Top, Data from a slice preexposed in control aCSF. Bottom, Slice preexposed with D-Asp. C, Summary data of SCR frequency in control and D-Asp preexposed slices from barrel cortex, CA1 hippocampus, and VB thalamus illustrating their sensitivity to AP5. D, Patch-clamped neuron in the presence of TTX with pipette containing Fluo-4. The images on the left illustrate fluorescence before and at peak of the SCR. Traces show a spontaneous recorded SIC (top) and elicited fluorescence SCR (bottom). E, Patched neuron with pipette containing Fluo-4 recorded in current-clamp mode. Traces show a spontaneous burst (top) and elicited fluorescence calcium signal (bottom). F, Spontaneous SCRs from single-neuron patch-clamp recordings (top, blue traces) normalized to amplitude to illustrate kinetics. Middle traces (green) show example spontaneous SCRs from Fluo-4 AM-loaded slices in TTX. Gray lines represent individual SCRs; colored lines, the mean. Bottom traces show the mean traces from the two conditions superimposed with symbols illustrating SEM. G, Top trace shows responses in a patch-clamped neuron to injection of recorded SIC waveforms. Middle trace shows a neuron response to the SICs in current-clamp mode, with expanded action potentials in numbered responses to the left. Red bottom trace shows fluorescence changes during the SIC evoked events. H, Nest of calcium responses normalized to peak amplitude generated by injected SICs (red) and generated by spontaneous SICs (blue) with averaged comparison to the right.