Fig. 2: Junctin variant transgenic zebrafish exhibit heat- and caffeine-induced motor defects. | Nature Communications

Fig. 2: Junctin variant transgenic zebrafish exhibit heat- and caffeine-induced motor defects.

From: Variants in ASPH cause exertional heat illness and are associated with malignant hyperthermia susceptibility

Fig. 2

a Immunolabeling of myofibers isolated from transgenic (Tg)-WT, Tg-V54A and Tg-K88T zebrafish with anti-mCherry and anti-RyR1antibodies. Magenta: mCherry, green: RyR1, scale bar: 5 μm. b Representative electron microscopy images from muscle of non-Tg, Tg-WT, Tg-V54A, and Tg-K88T zebrafish. Scale bar: 500 nm. c An overview of the workflow of the Heat + Caffeine assay. d Swimming ability of junctin mutant transgenic fish is altered with heat + caffeine exposure. Fish were tested without treatment (left panel) or with 1 mM caffeine at 34 °C for 1 h (right panel). Distance traveled was measured and normalized to the mean of non-Tg tested from the same day. Each group consists of 60 fish, which were obtained from three independent experiments with 20 fish per group. Error bars represent SEM. e Synergistic effect of heat and caffeine on the swimming ability of fish injected with junctin mRNA. Scatter plots display the total distance traveled normalized to the mean of non-injected measured on the same day. Data from non-injected (n = 100) and the other groups (n = 60) were from independent experiments with 20 animals per group at a time. The treatment conditions for each are no treatment (the leftmost panel), 1 mM caffeine at 28.5 °C for 1 h (the second panel from the left), at 34 °C for 1 h (the second panel from the right), and 1 mM caffeine at 34 °C for 1 h (the rightmost panel). Error bars represent SEM. Statistical analysis for (d, e) by one-way ANOVA followed by Tukey’s (d) and Dunnett’s (e) multiple comparisons tests where *p < 0.05, ***p < 0.001, ****p < 0.0001. Source data are provided as a Source data file.

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