TRANSCRIPTOMIC ANALYSIS OF ZINC OXIDE NANOPARTICLE TOXICITY ON THE REGENERATION ABILITY OF ZEBRAFISH (DANIO RERIO) CAUDAL FIN

• Main Author: Angellia, Vonny
• Format: Final Project
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/76521
• Institution: Institut Teknologi Bandung

The utilization of zinc oxide nanoparticles (ZnO NPs) has experienced a significant increase due to their various functions, including drug delivery systems, biosensors, and nanomedicine. However, ZnO NPs have the potential to induce toxic effects on cells and tissues, such as excessive production of reactive oxygen species (ROS), DNA damage, and chronic inflammation if not used optimally. The precise negative impacts caused by ZnO NPs are still not fully understood, and in-depth research in this area remains limited to date. Therefore, this study aims to investigate the influence of ZnO NP exposure on test animals. The study evaluates the effects of ZnO NPs on the regenerative capacity of the caudal fin in zebrafish (Danio rerio) at the transcriptomic level. Zebrafish were observed for caudal fin growth after being exposed for 30 minutes to E3 1X medium as the negative control group and ZnO NPs at a concentration of 72.59 ppm (LC50) as the treatment group, with each sample group consisting of 2 technical replicates. The surface area and blastema growth length were observed 96 hours post-amputation (hpa) using an optical microscope and analyzed with Image-J. RNA sequencing of the blastema was conducted using Illumina NextSeq PE150 Sequencing. Pre-processing of the total blastema RNA sequences was performed using FastQC v.11.9 and Cutadapt v.2.8. Gene mapping and counting were conducted with Rsubread v.2.12.2. Differential expression analysis was performed using EdgeR (Log2FoldChange>|1|, pvalue< 0.05). Enrichment analysis to explore gene ontologies was carried out using DAVID, iDEP, and REACTOME. Visual observations showed that zebrafish in the treatment group with ZnO NPs exhibited smaller blastema growth compared to zebrafish in the negative control group (p<0.05). The results of differential gene expression analysis identified 333 genes significantly affected by ZnO NP LC50, with 83 genes showing increased expression and 250 genes showing decreased expression. Administration of ZnO NP LC50 to zebrafish caudal fin regeneration significantly influenced zinc ion binding and transport processes regulated by the fthl28 gene, immune response regulated by the nlrc11 and prg4a genes, as well as lipid homeostasis regulated by the hmgcra, msmo1, and cyp7a1 genes. This research is expected to provide fundamental information for finding solutions and alternatives in the utilization of zinc oxide nanoparticles.