Schematic representation of the assay tank for locomotor assessment retrieved from Noldus EthoVision XT software version 11.5. Solid line: arena area; dashed line: imaginary quadrant divisions; Calib: calibration.

Chronological sequence of the experimental procedure. Locomotor assessment (indicated by a video icon) was conducted at three specific trial points: prior to injection, 24 h post-injection, and 48 h post-injection. Following the initial locomotor assessment, MPTP/saline was administered (indicated by a syringe icon). All fish were sacrificed immediately after the third locomotor assessment (indicated by a skull icon).

Effect of MPTP neurotoxin on mitochondria and HMGB1-associated genes in adult zebrafish. Whole RT-qPCR analysis for (A) mitochondrial genes; fis1, pink1, and park2, (B) HMGB1/neuroinflammatory genes; hmgb1, tlr4, and nfkb (n = 5 whole brains from each respective group). Bars represent the mean ± SEM. * p < 0.05, *** p < 0.005.

MPTP effect on adult zebrafish swimming behavior. (A–D) Analysis of total distance, average speed, freezing duration, and crossing frequency for MPTP-injected zebrafish and controls. Representative path images showing the movement of (E–G) controls, and (H–J) MPTP-injected zebrafish throughout the behavioral test period. Baseline, 24 h, and 48 h represent movement before MPTP/vehicle administration, 24 h, and 48 h after administration, respectively (n = 10 for both groups). Bars represent the mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.005, **** p < 0.001.

A gene network connecting mitophagy-associated and HMGB1-mediated inflammatory genes was generated with Pathway Studio^® (https://mammalcedfx.pathwaystudio.com/). Each link is supported by at least one published reference. Description on color-coded arrows can be found in the legends located in the bottom-right corner.