Knockdown of glulb induces apoptosis in zebrafish hair cells, and the supplementation of glutamine fails to rescue the observed phenotype. (A) The zebrafish embryos treated with glulb-MO and the control zebrafish were subjected to TUNEL staining, which revealed the presence of apoptotic signals, specifically in the hair cells of the glulb-MO-treated zebrafish. (B) The percentage of hair cells displaying apoptotic signals was evaluated in control zebrafish embryos and glulb-MO-injected zebrafish. TUNEL+/GFP+ denotes hair cells exhibiting apoptotic signals, while TUNEL-/GFP+ represents hair cells lacking apoptotic signals. Chi-squared test: ****, p < 0.0001. (C) The phenotype of hair cell clusters in whole embryonic zebrafish was visualized using confocal laser microscopy. Significantly reduced hair cell clusters were observed in glulb-MO-treated zebrafish compared to Ctrl zebrafish, and co-injection of glulb-MO and glutamine failed to rescue the reduction in hair cell clusters. (D) Similarly, a decreased number of hair cells was observed in the inner ear of zebrafish embryos treated with glulb-MO compared to control zebrafish, and co-injection of glulb-MO and glutamine failed to rescue the phenotype. Lateral crista (LC), anterior crista (AC) and posterior crista (PC). (E) The number of hair cells exhibited a significant reduction statistically, with no substantial rescue observed following glutamine supplementation. One-way ANOVA: ****, p < 0.0001; **, p < 0.01, ns means no significant.
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