Screening and Stability Analysis of Housekeeping Genes for Transcriptomic Studies in Danio rerio (Zebrafish) at Early Developmental Stages

• Published in: Molecular genetics, microbiology and virology microbiology and virology, 2024-06, Vol.39 (2), p.131-136
• Main Authors: Partevian, S. A., Safina, D. R., Rybolovlev, I. N., Rudenok, M. M., Kostrov, S. V., Shadrina, M. I., Slominsky, P. A., Alieva, A. Kh
• Format: Article
• Language: English
• Subjects: Biomedical and Life Sciences; Danio rerio; Developmental stages; Embryos; Experimental Papers; Gene expression; Hatching; Life Sciences; Microbiology; Molecular Medicine; Transcriptomics
• ISSN: 0891-4168; 1934-841X
• DOI: 10.3103/S0891416824700150

The zebrafish ( Danio rerio ) is one of the promising objects for modeling pathological conditions. A large number of such works are carried out at the transcriptomic level. Transcriptomic studies require the use of stable housekeeping genes (HKGs), and so we selected candidate HKGs and analyzed their stability in D. rerio under normal conditions. The work was carried out using D. rerio of the AB line whose embryos were sampled at three time points: the hatching stage (2 days postfertilization, dpf), and early larval stages (5 and 9 dpf). Analysis of HKG expression levels was performed using real-time polymerase chain reaction. Further analysis of the obtained Ct results was carried out in the RefFinder program. Different HKGs are suitable for assessing changes in gene expression at different stages of D. rerio development. When assessing changes in gene expression at the hatching stage (2 dpf), the optimal pair of HKGs is aars1 and polr2f ; for the early larval stage (5 dpf), it is aars1 and psmd6 ; and for 9 dpf, it is bcat2 and actb1 . During transcriptomic studies conducted at several developmental stages simultaneously, it is necessary to use a single selected pair of HKGs for all the studied stages of zebrafish development: aars1 and lsm12b . According to the data obtained, aars1 and lsm12b are the most stable HKGs for all the early developmental stages of D. rerio studied. At the same time, we first showed that aars1 is the most stable HKG for the early stages of the development. The second most stable HKG is lsm12b . Our data on the stability of the lsm12b gene are consistent with previously published data.