Molecular and structural analyses of voltage-dependent anion channel 2 and its anti-apoptotic function in stress and pollutant resistance in Pacific abalone.

This study aimed to identify voltage-dependent anion channel 2 (Hdh-VDAC2) and determine its functional role in response to acute thermal stress, H2O2-induced stress, heavy metal toxicity, bacterial and viral infections, and during metamorphosis. Structural analysis confirmed that Hdh-VDAC2 is a pore-forming β-barrel protein. Molecular docking further confirmed the protein-protein interactions of Hdh-VDAC2 with Hdh-BAX, Hdh-caspase 3, and Hdh-BCL2. In the Hdh-VDAC2-inhibited hemocytes (HCY), apoptotic genes (Hdh-caspase-3 and Hdh-BAX) exhibited elevated mRNA expression, while the anti-apoptotic gene (Hdh-BCL2) was downregulated. Further, fluorescent techniques confirmed excessive reactive oxygen species (ROS) production, lower cell viability, elevated caspase 3 activity, and increased DNA fragmentation in Hdh-VDAC2-inhibited HCY, indicating an anti-apoptotic role of Hdh-VDAC2 in Pacific abalone. Transcriptomic analysis revealed differential expression patterns, with upregulation in the digestive gland (DG) and downregulation in the gill (GIL) and HCY when comparing heat-tolerant (HT) versus heat-sensitive (HS) abalone groups. Additionally, both cold and heat stresses induced Hdh-VDAC2 expression. Other environmental factors including H2O2, cadmium, bacteria, and viruses, were also shown to induce Hdh-VDAC2 mRNA expression in the GIL and DG of Pacific abalone. During metamorphosis, the blastula (BLS) stage exhibited higher Hdh-VDAC2 mRNA expression. These findings suggest that Hdh-VDAC2 plays a crucial anti-apoptotic role and may be a biomarker for summer mortality in Pacific abalone.