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. Author manuscript; available in PMC: 2022 Oct 28.
Published in final edited form as: Cell. 2021 Oct 4;184(22):5622–5634.e25. doi: 10.1016/j.cell.2021.09.013

Fig. 3: GRP depolarizes and increases intracellular Ca2+ in cortical VIP cells.

Fig. 3:

A, Schematic of whole-cell recording of ACx VIP cell used to examine effects of GRP.

B, Two exemplary VIP cells responding with bursts (top) or depolarization (bottom) following 2 min bath application of GRP (300 nM) in the presence of NBQX, CPP, gabazine, CGP. Inset: magnification of first bursts indicated by an asterisk.

C, Depolarization of VIP cells upon application of indicated concentrations of GRP. Mean ± SEM. 6-10 cells per group. Comparison 0 vs. 300 nM GRP: t-test for unequal variance: t(12.52)=3.76, p=0.01. Other comparisons n.s. Bath contains NBQX, CPP, gabazine, CGP.

D, Representative firing patterns of ACx VIP, SST, PVALB and pyramidal (Pyr) cells upon −200 pA current injection (bottom), at AP threshold (middle), and at maximal firing rate (top).

E, Average time course (left) and amplitude (right) of the membrane potential changes in each indicated cell type in ACx following GRP application. Bath contains NBQX, CPP, gabazine, CGP, TTX. Mean ± SEM. n=10 VIP, SST, Pyr cells and 15 PVALB cells. Comparison to VIP cells (Bonferroni-corrected t-test): SST: t(18)=−5.27, p<0.001; PVALB: t(23)=−3.83, p<0.001; Pyr: t(18)=−4.87, p<0.001.

F, Design of plasmid for Cre-dependent stoichiometric expression of GCaMP and mBeRFP for imaging of Ca2+ entry and detection of infected cells, respectively.

G, Injection of AAV DIO-GCaMP-P2A-mBeRFP into ACx of male VipCre mice (left) and epifluorescent GCaMP imaging (right) in acute slices before (top) and after (bottom) GRP bath application, in the presence of NBQX, CPP, gabazine and CGP.

H, Heatmap of fluorescence changes (expressed relative to fluorescence following KCl application, ΔF/FKCl) across all imaged VIP cells in an exemplary acute ACx slice.

I, GCaMP fluorescence changes with or without TTX in two exemplary VIP cells (top) or across all recorded cells (bottom) in the presence of NBQX, CPP, gabazine and CGP. Mean ± SEM, Mann-Whitney U test: U=10178, p<0.0001. n=218 (−TTX) and 179 (+TTX) cells in 7 and 6 slices respectively.

See also Fig. S3.