Journal: Cell Stress & Chaperones

Article Title: The role of Atp2a2 -mediated calcium imbalance and endoplasmic reticulum stress in hydrocortisone-induced neurotoxicity

doi: 10.1016/j.cstres.2025.100112

Figure Lengend Snippet: Effects of RU486 and S2A1 on calcium homeostasis and mitochondrial membrane potential in HC-treated zebrafish larvae. (a) Fluorescence staining images of zebrafish larvae (120 hpf) in different treatment groups, focusing on the brain region: green fluorescence represents cytoplasmic Ca²⁺ labeled by Mag-Fluo-4 AM, red fluorescence represents mitochondrial Ca²⁺ labeled by Rhod-2, and the merged image shows the colocalization of the two. The merged image illustrates the colocalization of the two signals. (b) Quantitative analysis of the fluorescence intensities of Mag-Fluo 4 AM and Rhod-2 (n = 3 replicates, N = 3 biological replicates, 15 embryos per replicate. Data are presented as mean ± SEM, * P < 0.05, ** P < 0.01, *** P < 0.001). (c) Representative images of zebrafish larvae stained with JC-1 to assess mitochondrial membrane potential (ΔΨm). Red fluorescence is the aggregated state of JC-1 (normal membrane potential), and green fluorescence is the monomer of JC-1 (membrane potential depolarization), with merged images representing the balance of the two signals. (d) Quantification of the JC-1 red/green fluorescence ratio, reflecting changes in mitochondrial membrane potential across different groups (n = 3 replicates, N = 3 biological replicates, 15 embryos per replicate; data are presented as mean ± SEM, * P < 0.05, ** P < 0.01, *** P < 0.001).

Article Snippet: To validate the ROS detection system, a positive control group was included: 15 normally developing 120 hpf larvae were treated with 1 mM H2O2 in E3 medium for 4 h. All larvae were then incubated in 40 μM DCFH-DA (MCE, USA) working solution at 28 °C in the dark for 1 h. After three washes in E3 medium, larvae were anesthetized with 0.03% MS-222 and imaged using a confocal microscope.

Techniques: Membrane, Fluorescence, Staining, Labeling

Journal: Cell Stress & Chaperones

Article Title: The role of Atp2a2 -mediated calcium imbalance and endoplasmic reticulum stress in hydrocortisone-induced neurotoxicity

doi: 10.1016/j.cstres.2025.100112

Figure Lengend Snippet: Effects of hydrocortisone on neuronal fluorescence signals and locomotor activity in zebrafish larvae. (a) Fluorescence imaging of transgenic zebrafish larvae (120 hpf). HuC-labeled neurons (green fluorescence) demonstrate the distribution and fluorescence intensity of neurons in the brain and spinal cord across different groups. (b) Quantification of neuronal fluorescence intensity. The y-axis represents fluorescence intensity. n = 3 replicates, N = 3 biological replicates, 15 embryos per replicate. Data are presented as mean ± SEM, * P < 0.05, ** P < 0.01, *** P < 0.001. (c) Representative behavioral heatmap of zebrafish larvae. The movement trajectories and activity distribution of juvenile fish, with color intensity reflecting movement frequency (red indicates high-frequency activity zones, and blue indicates low-frequency activity zones). (d) Quantification of locomotor speed in zebrafish larvae (n = 3 replicates, N = 3 biological replicates, 6 embryos per replicate). The y-axis represents swimming speed (mm/s), and data are presented as mean ± SEM. Significance levels: * P < 0.05, *** P < 0.001.

Article Snippet: To validate the ROS detection system, a positive control group was included: 15 normally developing 120 hpf larvae were treated with 1 mM H2O2 in E3 medium for 4 h. All larvae were then incubated in 40 μM DCFH-DA (MCE, USA) working solution at 28 °C in the dark for 1 h. After three washes in E3 medium, larvae were anesthetized with 0.03% MS-222 and imaged using a confocal microscope.

Techniques: Fluorescence, Activity Assay, Imaging, Transgenic Assay, Labeling