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. 2003 Dec 17;23(37):11554–11567. doi: 10.1523/JNEUROSCI.23-37-11554.2003

Figure 2.

Figure 2.

Whole-cell recordings and fura-2 recordings of Ca2+ accumulation for the various fluorescently tagged nicotinic receptor constructs expressed in HEK293T cells. ACh (300 μm) was applied every 60 sec (duration 50 msec). A, Exemplar ACh-induced currents of each of the fluorescently tagged chimeras. Currents were inhibited with continuous coapplication of the competitive antagonist DHβE (10 μm) and recovered during washing. B, Peak current response with 300 μm ACh of the fluorescently tagged and WT α4β2 receptors. Numbers represent mean ± SEM. Asterisks represent a significant difference (p < 0.05) as compared with WT. Ratiometric measurements (F340/F380) of fura-2 AM loaded in HEK293T cells show that intracellular calcium rises during a 5 sec application of 300 μm ACh for WT α4β2, α4-YFP-M β2, and α4 β2-CFP-M (C). Application of 300 μm ACh to cells expressing α4-YFP-N1 β2 and α4 β2-CFP-C showed no change in levels of intracellular calcium. Each trace represents a single cell. D, Box plot of peak calcium fluxes for each of the fluorescently labeled constructs. Bottom, middle, and top line of the box represents 25%, median, and 75%, respectively. Bottom error bar represents 5%, and the top error bar represents 95%. Asterisks represent a significant difference (p < 0.05) as compared with WT.