ZFIN Image: Linsley et al., 2021, Fig. 4

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Figure Caption

Fig. 4 Comparison of GEDI variants to detect neuronal death.

A–C Representative images of rat primary cortical neurons expressing RGEDI-P2a-3xBFP (A), mRuby-P2a-GCaMP6f (B), or RGEDI-P2a-EGFP (C), before, 5 min and 10 min after exposure to NaN₃. D Quantification of the peak signal and response rate of signal increase (τ) from fitted nonlinear regressions of increases in fluorescence signals over time from variants of the GEDI biosensor: RGEDI-P2a-3xBFP (n = 23), mRuby-P2a-GCaMP6f (n = 52), RGEDI-P2a-EGFP (n = 41), GC150-P2a-mApple, RGEDI-NLS-P2a-EGFP-NLS, and GC150-NLS-P2a-mApple-NLS (n = 40), compared to EGFP-P2a-mApple (n = 18). Error bars represent SE, ANOVA Tukey’s ****p < 0.0001; ns not significant, n values represent individual neurons sampled across at least three independent wells. E Relative fluorescence of GECI’s GCaMP6f⁷⁹ and RCEPIA³⁶/RGEDI and GCaMP150ER³⁷/GC150 across Ca²⁺ concentrations modeled from previously reported Hill coefficients and K_d values. F Representative images of rat primary cortical neurons expressing GC150-P2a-mApple, G GC150-NLS-P2a-mApple-NLS, and H RGEDI-NLS-P2a-EGFP-NLS before, 5 min, and 10 min after exposure to NaN₃. Error bars represent SEM. Scale = 25 μm. Experiments in A–C and F–H were repeated at least 18 times with similar results.

Acknowledgments

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