Breaking Barriers: A Reliable Method for RNA Extraction in Zebrafish Neurodegenerative Research

Zebrafish (<em>Danio rerio</em>) is a vertebrate with unique characteristics, making it an excellent biomedical research animal model. One research area involves investigating changes in gene expression during neurodegenerative processes, aging-related changes, and behavioral studies in zebrafish. To achieve this, high-quality and quantity RNA must be extracted from zebrafish. This study aims to develop an optimized RNA extraction method from zebrafish larvae brains. After rearing and maintaining the zebrafish in suitable conditions, the larvae were separated, and their brains were extracted. Using an optimized TRIzol-based method, RNA was extracted from the zebrafish larvae's brains. For comparison, we also employed commercial kits and Trizol for RNA extraction. The quantity and quality of the extracted RNA were measured using NanoDrop, gel electrophoresis, and real-time PCR. We selected five genes: <em>gapdh</em> and <em>beta-actin </em>as housekeeping genes and three other genes expressed in the brain and nervous system (<em>bdnf, chd8, and lrp6</em>). According to NanoDrop results, all samples were purified with minimal protein and polysaccharide contamination, and the 260/280 ratio and 260/230 ratio fell within the standard range of 1.9-2.2. The amount of extracted RNA significantly increased compared to other methods (P value < 0.0001), and all the studied genes exhibited high expression. RNA extraction demonstrated both high quality and quantity. In summary, we have developed an optimized method for RNA extraction from the brains of zebrafish larvae.