Abstract
Early life stress is key in development and behaviour and may have long-term effects, as it represents a window of increased vulnerability. In our study we evaluated two stressors, temperature and photoperiod —alone and combined—, in zebrafish ( Danio rerio ) larvae up to 120 h post-fertilisation. The objective of this study was to investigate the effects of non-lethal environmental stressors and their potential synergistic effects until completion of organogenesis integrating molecular, cellular and behavioural analyses. Larvae were subjected to 4 experimental conditions: “control” (C) (14 h light: 10 h darkness; T = 27 ± 1 °C); “heat shock” (HS) (14 h light: 10 h darkness; T = 34 °C); “darkness” (D) (24 h darkness; T = 27 ± 1 °C) and “stressed” (S + ) (24 h darkness; T = 34 °C). Our results confirmed phenotypic, behavioural and molecular alterations. The S + larvae showed a lower survival curve, lower regeneration, a higher number of malformations, predominantly those related to the swim bladder, lower motor activity and a dysregulation of miR29a and clocka , hspa9 , hspa5 and cxcr4b genes. The results revealed a similar number of Ddx4 + cells between groups but non-canonical migration patterns in the S + group. This integrated approach offers new insights into the molecular mechanisms underlying the stress response during embryogenesis and provides a new perspective on the interaction between temperature and light on early-stage development.