Effects of the Tripeptide treatment before pentylenetetrazol (PTZ)-induced seizures on seizure-like behaviors and swimming activity.

Animals were exposed to the Tripeptide for 24 hours before to Pentylenetetrazol (PTZ)- induced seizures. Groups included: PTZ control (n=10); Tripeptide treatment at 25 µg/mL (T25; n=10); 10 µg/ml (T10; n=10); 5 µg/ml (T5; n=10). Animals were video-recorded during a 10-minute exposure to PTZ (15mM). (A) The number of seizures was scored by visual analysis twice per larva, with loss of posture serving as the criterion for a complete seizure event; (B) Distance traveled and (C) velocity for each larva were quantified using EthoVision XT locomotion tracking software (Noldus, Wageningen, The Netherlands). Data are presented as mean ± SEM. Statistical analyses were performed using one-way ANOVA, followed by the Bonferroni correction for multiple comparisons. Differences were considered significant at p < 0.05.

Effects of CX2 treatment before pentylenetetrazol (PTZ)-induced seizures on the number of seizure-like behaviors and swimming activity.

Animals exposed to CX2 for 24 hours before Pentylenetetrazol (PTZ)- induced seizures. Groups included: PTZ control (n=10); CX2 treatment at 0.5 µM (T0.5; n = 9), 0.1 µM (T0.1; n = 10), and 0.05 µM (T0.05; n = 10). Animals were video-recorded during a 10-minute exposure to PTZ (15 mM). (A) The number of seizures was scored visually twice per larva, with loss of posture as the criterion for a complete seizure event. (B) Distance traveled and (C) velocity of each larva were quantified using EthoVision XT locomotion tracking software (Noldus, Wageningen, The Netherlands). Data are expressed as mean ± SEM. Statistical analyses were conducted using one-way ANOVA followed by Bonferroni correction for multiple comparisons. Differences were considered significant at p < 0.05.

Expression levels of il1b, casp3a, casp9, baxa, bcl2a, and c-fos genes in zebrafish brain after pentylenetetrazol (PTZ)-induced seizures.

Each treatment group was pre-exposed to the Tripeptide for 24 h, followed by 15 mM PTZ exposure for 20 min. Control and PTZ groups underwent identical handling procedures but were exposed to water (n = 5 per group). Data are presented as mean ± SEM. Statistical analyses were performed with the one-way ANOVA, followed by Bonferroni’s test for multiple comparisons. Differences were considered significant at p<0.05. Significance is indicated as follows: *p≤0.05; **p≤0.01; ****p≤0.0001. Groups: Control (Control), PTZ (pentylenetetrazol group), T25 (25 µg/mL treatment), T10 (10 µg/mL treatment), T5 (5 µg/mL treatment).

Expression levels of il1b, il6, tnfa, cox1, cox2a and c-fos genes in zebrafish brain after pentylenetetrazol (PTZ)-induced seizures.

Each treatment group was pre-exposed to CX2 for 24 h, followed by exposure to 15 mM PTZ for 20 min. Control and PTZ groups underwent identical procedures but were exposed to water only (n = 5 per group). Data are presented as mean ± SEM. Statistical analyses were performed with the one-way ANOVA, followed by Bonferroni’s multiple comparisons test. Differences were significant at p<0.05. Significance indicators: *p≤0.05; **p≤0.01; ***p≤0.001; ****p≤0.0001. Groups: Control (Control), PTZ (pentylenetetrazol group), T0.5 (0.5 µM treatment), T0.1 (0.1 µM treatment), T0.05 (0.05 µM treatment).

Time course of CX2 (8ARG-Cx43p, corresponding to the C-terminal domain sequence of Cx43) crossing the blood-brain barrier (BBB) efficiently in a 7-day-old zebrafish larva.

The carboxytetramethylrhodamine (TAMRA)-labeled peptide showed fluorescent signal detection (indicated by red color) in the brain and throughout the body at 1 h and 6 h post-treatment, as observed by fluorescence microscopy.