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Long-term effects of early-life thermal fluctuations on the cellular stress response and CTmax of zebrafish, Danio rerio

Preprint Created on 08 Jun 2026 bioRxiv

Temperatures in aquatic ecosystems have been affected by anthropogenic activities such as agricultural and industrial water use, and climate change caused by greenhouse gas emissions. Changes in water temperature directly affect the cellular and organismal physiology of fishes because most fishes are ectotherms. These effects can take different forms, such as increased cellular stress and reactive oxygen species (ROS) production. However, the type and magnitude of response depend on the duration and the frequency of the exposure to elevated water temperature. In this study, we investigated the long-term effects of exposure to daily thermal fluctuations occurring during early-life stages of zebrafish, Danio rerio, on gene expression and CTmax in later developmental stages. To do so, wild-type zebrafish were exposed daily to a + 5 {degrees}C fluctuation in temperature from ambient (28 {degrees}C) to 33 {degrees}C over the first 30 days post fertilization (dpf), before being held until 90 dpf at ambient temperature. The fish that experienced daily thermal fluctuation were compared to a control group that was kept at 28 {degrees}C throughout the experiment and sampled at the same timepoints. Samples were collected at 18 (larval), 30, 60 (juvenile), and 90 (adult) dpf to study the expression of heat shock proteins and oxidative stress genes. The thermotolerance of fish was tested using CTmax trials at 60 and 90 dpf. Daily thermal fluctuation over the first 30 dpf led to a significant increase in the expression of hsp47, gstp1a, sod1, and sod2 genes at 60 dpf, and hsp47, hsp90aa1, hsp90ab1, cat, glulb, gstp1a, sod1, and sod2 genes at 90 dpf. The only significant increase detected during the larval stage was glulb at 18 dpf. Fish that experienced thermal fluctuation also had a higher CTmax at 60 dpf, but this increased thermotolerance significantly decreased from 60 to 90 dpf, where it was not different between treatments. Overall, our study demonstrates that early-life thermal stress increased cellular stress responses and thermotolerance in zebrafish into later ontological stages.

Haghighi, H., Lindsey, B. W., Jeffries, K. M.

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