Fig. 5: Spontaneous and evoked calcium activity analysis of 2D cortical cultures at day 70.
From: Novel fragile X syndrome 2D and 3D brain models based on human isogenic FMRP-KO iPSCs
A Calcium traces as a function of ΔF/F0 of FAST (top) and SLOW (bottom) active cells. B Representative calcium trace (insert from panel A) showing extracted functional properties: rising time (a), amplitude (b), and event frequency (c). C FAST active cells with respect to total cells in both FMRP-KO and FMRP-WT populations (n = 14 FOVs for genotype; 144 ± 13 total cells in WT, 28 ± 6 active cells in WT; 168 ± 8 total cells in KO, 34 ± 5 active cells in KO; rising time 1.3 ± 0.1 sec in WT and 1.3 ± 0.03 sec in KO; differentiation batches=1). D SLOW active cells with respect to total cells in both FMRP-KO and FMRP-WT populations (n = 14 FOVs for genotype; 144 ± 13 total cells in WT, 56 ± 11 active cells in WT; 168 ± 8 total cells in KO, 113 ± 9 active cells in KO; rising time 5.2 ± 0.6 sec in WT and 5.1 ± 0.17 sec in KO; differentiation batches = 1). E Graph plots reporting the functional properties of FAST active cells: amplitude (left, n = 14 FOVs, *p < 0.05 KO vs WT, MW test), frequency and relative cumulative distribution (middle, n = 14 FOVs, 0.035 Hz in WT and 0.06 Hz in KO, *p < 0.05 KO vs WT) and synchronicity (right) of spontaneous calcium events of FMRP-WT and FMRP-KO fast cells (n =14, 0.43 ± 0.09 in WT and 0.41 ± 0.06 in KO not significant). F Graph plots reporting the functional properties of SLOW active cells amplitude (left, n = 14 FOVs, *p < 0.05 KO vs WT, MW test), frequency and relative cumulative distribution (middle, n = 14 FOVs, 0.025 Hz in WT and 0.025 Hz in KO, not significant) and synchronicity (right) of spontaneous calcium events of FMRP-WT and FMRP-KO slow cells (n = 14, 0.51 ± 0.05 in WT and 0.6 ± 0.03 in KO, not significant). G Representative FOV showing a puffer pipette for local Glutamate and GABA application with a cartoon showing the experimental approach. H Glutamatergic calcium transient response of FMRP-KO and FMRP-WT cortical neurons after puffer applications (n = 5 FOVs, 23 ± 4.7 total cells and 17 ± 4 active cells in WT; n = 4 FOVs, 22 ± 2 total cells and 18 ± 1 active cells in KO; **p < 0.01 KO vs WT; differentiation batches = 1). I GABAergic calcium transient response of FMRP-KO and FMRP-WT cortical neurons after puffer applications (n = 5 FOVs, 22.6 ± 3.5 total cells and 7 ± 1.8 active cells in WT; n = 5 FOVs, 14.6 ± 1.7 total cells and 11.4 ± 2 active cells in KO, **p < 0.01 KO vs WT; differentiation batches = 1).
