Identification and Characterization of a G Protein-binding Cluster in alpha7 Nicotinic Acetylcholine Receptors 1.0.0

In α7345–348A nAChR expressing cells, nifedipine had no effect on the peak or the duration of the calcium transient (peak: 957.00% ΔF/Fθ ± 252.2%; AUC: 333.33% ΔF/Fθ2 × s ± 91.53%) relative to choline treatment alone (Fig. 5, A–C). The findings suggest that choline-induced calcium responses in PC12 cells involve the activity of VGCC (37, 38). PubMed:26088141

PC12 cells transfected with α7345–348A showed a reduction in choline-mediated calcium responses. PubMed:26088141

In α7345–348A nAChR expressing cells, nifedipine had no effect on the peak or the duration of the calcium transient (peak: 957.00% ΔF/Fθ ± 252.2%; AUC: 333.33% ΔF/Fθ2 × s ± 91.53%) relative to choline treatment alone (Fig. 5, A–C). The findings suggest that choline-induced calcium responses in PC12 cells involve the activity of VGCC (37, 38). PubMed:26088141

Treatment of PC12 cells with 10 mM choline was associated with a translocation of PH-mCherry from the cell surface as determined by the presence of the fluorescence signal within 1 μm of the edge of the cell into the cytosol of the GC (Fig. 6, A and B). Pre-treatment of cells with SP abolished this translocation (Fig. 6B). PubMed:26088141

Sequential imaging of PH-mCherry and GCaMP5G confirms that choline promotes a rise in intracellular calcium and PH-mCherry translocation in the same cellular compartment (Fig. 6, B and C). Cytoplasmic translocation of PH-mCherry occurred on a slower time scale (40 s after choline application) than peak calcium responses (∼1 s after choline application). These kinetics are consistent with the translocation of the PH domain sensor in the cell (20, 29). PubMed:26088141