Chemical mutagenesis substances have been studied in various ways, such as body color change by inducing mutations for the purpose of developing application technologies for the aquarium fish industry. In this study, three chemical mutants of three chemical mutagenic substances were immersed in concentrations of 1, 2, 3 and 5 mM, and then treated to find an appropriate concentration. As a result, the optimal concentration of zebrafish was confirmed at 2 mM. As a result, they acquired the F1 generation and three chemical mutagenic substances, 1-ethyl-1-nitrosourea (ENU), colchicine and hydroxylamine, were used to investigate the effect on zebrafish (Danio rerio) immunity (lysozyme), growth-related (ghra) and nerve-related (nr4a2b) genes.
The zebrafish breeding water was immersed for 1 hour 3 times for 6 days at a concentration of 2 mM of each chemical mutagenic substances. And then after7 days, the brain, liver and kidney were extracted and total RNA was extracted. After total RNA was synthesized into cDNA, changes in genes were analyzed using quantitative PCR.
After immersion in 2 mM concentration, ghra did not show a sex difference in ENU and hydroxylamine in gene expression analysis, but showed more than double the expression in males of colchicine compared to females of zebrafish. In contrast to growth, lysozyme, an immune-related gene, did not show a male and female difference in colchicine, and showed higher expression in females than in males in ENU and hydroxylamine. In addition, it showed a significant increase compared to the control. The nr4a2b gene expression showed similar to that of the control in F1 crossed with normal males and hydroxylamine females. Our results, ENU and hydroxylamine at 2 mM affect the cleavage, and colchicine can be male of zebrafish. And results related to immunity and growth related to mutant substances in previous studies have not been reported much, and it is judged that the results of this study can be used as basic data. And also, these results will compare and analyze the mutation changes in F2, the offspring of F1, following the previously conducted in zebrafish treated with mutation inducing chemicals and their progeny F1.