Figure 7.
Overexpression of TMBIM-2 in neurons preserves age-onset decline of aversive learning of C. elegans. (A) Correlation between normalized expression of Tmbim2 in the prefrontal cortex of human brains with Alzheimer’s disease at different ages. Expression values in A (n = 101 samples) are from the dataset GSE44772. (B) Correlation between normalized expression of Tmbim2 in mouse brains at different ages. Expression values are from the dataset GSE132040 (n = 38 samples). (C) Correlation between normalized expression of srh-142 positive neurons at different ages. Expression values are from Roux et al. (2023) (n = 147 cells). (D) Representative confocal photomicrograph of animals head expressing TMBIM-2 at different ages. The imaging used z-planes. Scale bar, 20 μm. (E) Quantitative analysis of TMBIM-2::GFP protein levels in ROI shown in D. n ≥ 8 worms. (F) Quantification of the amplitude of PM-GCaMP6f fluorescence intensity changes in expressed neuronal PM-GCaMP6f and ADF presynaptic vesicle marker worms with neuronal cox-5B knockdown at different ages. (G) Example of control and aversion phenotypes after exposure to pathogenic P. aeruginosa PA14 in WT, tmbim-2, tmbim-2p::tmbim-2::gfp and tmbim-2; tmbim-2p::tmbim-2::gfp animals at day 1. Scale bar, 250 μm. (H) Quantification of the percentage of the worm on PA14 at day 1 as shown in G. n = 2 independent experiments. (I) Quantification of the percentage of the worm on PA14 in WT and tmbim-2p::tmbim-2::gfp animals at day 5 as shown in G. n = 2 independent experiments. (J) Survival analyses of WT (solid blue); tmbim-2 (dashed blue); unc-119p::cox-5B HP (solid yellow) and unc-119p::cox-5B HP; tmbim-2 (dashed yellow) animals; n ≥ 100 worms. (K) Survival analysis of WT (blue) and tmbim-2p::tmbim-2::gfp (yellow); n ≥ 100 worms. (L) PA14 survival analyses of WT (black), tmbim-2 (yth26) (dashed blue), tmbim-2 (yth57) (dashed yellow), TMBIM-2 overexpression line1 (solid blue), and TMBIM-2 overexpression line2 (solid yellow) at day1 adult animals; n ≥ 100 worms. (M) PA14 survival analyses of WT (black), tmbim-2 (yth26) (dashed blue), tmbim-2 (yth57) (dashed yellow), TMBIM-2 overexpression line1 (solid blue), and TMBIM-2 overexpression line2 (solid yellow) at day5 adult animals; n ≥ 100 worms. ***P < 0.001, **P < 0.01, *P < 0.05, ns denotes P > 0.05 via unpaired two-tailed Student’s t test. Error bars, SEM. In A–C, we used Pearson’s correlation coefficient for statistical testing. In J–M, we used The Log-rank (Mantel-Cox) test for statistical analysis.

Overexpression of TMBIM-2 in neurons preserves age-onset decline of aversive learning of C. elegans. (A) Correlation between normalized expression of Tmbim2 in the prefrontal cortex of human brains with Alzheimer’s disease at different ages. Expression values in A (n = 101 samples) are from the dataset GSE44772. (B) Correlation between normalized expression of Tmbim2 in mouse brains at different ages. Expression values are from the dataset GSE132040 (n = 38 samples). (C) Correlation between normalized expression of srh-142 positive neurons at different ages. Expression values are from Roux et al. (2023) (n = 147 cells). (D) Representative confocal photomicrograph of animals head expressing TMBIM-2 at different ages. The imaging used z-planes. Scale bar, 20 μm. (E) Quantitative analysis of TMBIM-2::GFP protein levels in ROI shown in D. n ≥ 8 worms. (F) Quantification of the amplitude of PM-GCaMP6f fluorescence intensity changes in expressed neuronal PM-GCaMP6f and ADF presynaptic vesicle marker worms with neuronal cox-5B knockdown at different ages. (G) Example of control and aversion phenotypes after exposure to pathogenic P. aeruginosa PA14 in WT, tmbim-2, tmbim-2p::tmbim-2::gfp and tmbim-2; tmbim-2p::tmbim-2::gfp animals at day 1. Scale bar, 250 μm. (H) Quantification of the percentage of the worm on PA14 at day 1 as shown in G. n = 2 independent experiments. (I) Quantification of the percentage of the worm on PA14 in WT and tmbim-2p::tmbim-2::gfp animals at day 5 as shown in G. n = 2 independent experiments. (J) Survival analyses of WT (solid blue); tmbim-2 (dashed blue); unc-119p::cox-5B HP (solid yellow) and unc-119p::cox-5B HP; tmbim-2 (dashed yellow) animals; n ≥ 100 worms. (K) Survival analysis of WT (blue) and tmbim-2p::tmbim-2::gfp (yellow); n ≥ 100 worms. (L) PA14 survival analyses of WT (black), tmbim-2 (yth26) (dashed blue), tmbim-2 (yth57) (dashed yellow), TMBIM-2 overexpression line1 (solid blue), and TMBIM-2 overexpression line2 (solid yellow) at day1 adult animals; n ≥ 100 worms. (M) PA14 survival analyses of WT (black), tmbim-2 (yth26) (dashed blue), tmbim-2 (yth57) (dashed yellow), TMBIM-2 overexpression line1 (solid blue), and TMBIM-2 overexpression line2 (solid yellow) at day5 adult animals; n ≥ 100 worms. ***P < 0.001, **P < 0.01, *P < 0.05, ns denotes P > 0.05 via unpaired two-tailed Student’s t test. Error bars, SEM. In A–C, we used Pearson’s correlation coefficient for statistical testing. In J–M, we used The Log-rank (Mantel-Cox) test for statistical analysis.

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