Figure 8.

ChAc neurons and controls showed similar spontaneous and neurotransmitter-induced Ca²⁺ signaling. A, Representative traces of transient cytosolic Ca²⁺ changes induced by bath application of GABA (100 μm), acetylcholine (ACh, 100 μm), glutamate (Glut, 50 μm), glycine (Gly, 100 μm), and KCl (50 mm) in fura-2-loaded neurons from control iPSCs. Intracellular Ca²⁺ changes are presented as ratios of the fluorescence signals obtained at 340 and 380 nm (F₃₄₀/F₃₈₀). B, Summary of cytosolic Ca²⁺ response amplitudes normalized to the basal Ca²⁺ level of the investigated cells (n) showed no significant differences between ChAc neurons and healthy controls. C, A similar fraction of ChAc and control cells responded to neurotransmitter application obtained from experiments as shown in A. Note: GABA and glycine induced depolarizing Ca²⁺ signals in ∼20% of the cells. Therefore, the GABAergic and glycinergic synaptic transmission partly have an excitatory function in ChAc neurons and controls. D, Analyzing spontaneous Ca²⁺ transients revealed no significant difference in the percentage of spontaneously active cells (∼30%), Ca²⁺ current frequency, or amplitudes in ChAc neurons compared with controls. E, Genomic expression of voltage-gated calcium channel subunits analyzed by qRT-PCR did not differ significantly between ChAc neurons and healthy controls. Data represent means ± SEM (p < 0.05, Mann–Whitney test).