Thyroxine (T3)‐mediated regulation of early cardiac repair in a chemical‐induced hypoxia/reoxygenation model of adult zebrafish (<i>Danio rerio</i>)

Hypoxia-mediated cardiac tissue injury and its repair or regeneration are one of the major health management challenges globally. Unlike mammals, lower vertebrate species such as zebrafish (Danio rerio) represent a natural model to study cardiac injury, repair and regeneration. Thyroxine (T3) has been hypothesised to be one of the endocrine factors responsible for the evolutionary trade-off for acquiring endothermy and regenerative capability in higher vertebrates. However, the specific targets of T3 during cardiac repair are still obscure. In this study, cardiac injury was generated in adult zebrafish by acute anaemia-induced hypoxia/reoxygenation (H/R) in the presence or absence of exogenous T3 alone or along with 1-850 (inhibitor of T3 receptor) and iopanoic acid (IOA, blocker of T3 release), respectively. A microarray analysis showed that 10,226 gene expression changes in expression across all experimental groups, providing a comprehensive understanding of the cardiac transcriptome. Analysis of 11 candidate genes was conducted using qRT-PCR and the findings aligned with the microarray data. Histological assessment by Masson's trichrome staining and immunofluorescence studies also corroborated the microarray data. GO enrichment analysis showed noteworthy involvement of T3 in the modulation of genes involved in oxidative stress, cardiac fibrosis, energy metabolism, autophagy, apoptosis and regeneration during the initial repair phase (7 days) of H/R-damaged cardiac tissue. Overall, this is the first study that presents a holistic picture of cardiac repair and regeneration post H/R injury in zebrafish and the effect of T3 pre-treatment on it.