Preparation of polyclonal antibody targeting Optic atrophy 1 (OPA1) of chicken and the effect of naringenin on OPA1 expression in the liver of broilers under heat stress

The mitochondrial protein optic atrophy 1 (OPA1) belongs to the dynamin-related GTPase family and plays a crucial role in maintaining mitochondrial morphology and function. Heat stress disrupts mitochondrial function, thereby impairing cellular viability and causing damage to multiple organs. Naringenin, a flavanone, has been reported to exert protective effects against various stressors. However, direct studies examining the relationship between naringenin and heat stress remain limited, and the association between OPA1 and heat stress has been scarcely investigated. In addition, there is a lack of antibodies that specifically and accurately target the chicken OPA1 protein.In the present study, a recombinant expression plasmid, pET32a-OPA1, was constructed using prokaryotic expression technology to express the exogenous gene. The recombinant target protein was produced through an induced expression method. Subsequently, rabbit anti-chicken OPA1 serum was generated by immunizing rabbits with purified and concentrated recombinant OPA1 protein as the antigen. Enzyme-linked immunosorbent assay (ELISA) results demonstrated that the antibody titer exceeded 1:51,200. Western blotting combined with immunofluorescence analysis revealed that heat stress significantly reduced OPA1 protein levels in liver tissue, indicating a strong association between OPA1 and the heat stress response. Furthermore, naringenin treatment up-regulated OPA1 mRNA expression, fluorescence intensity, and protein levels. These findings provide preliminary insights into the regulatory effects of naringenin on mitochondrial OPA1 during heat stress conditions.