The Megalobrama amblycephala transferrin and transferrin receptor genes: Molecular cloning,characterization and expression during early development and after Aeromonas hydrophila infection

Both transferrin (TF) and transferrin receptor (TFR) play vital roles in iron homeostasis, which is essential for cellular growth and survival. Besides, TF and TFR are also reported to be involved in immune response against bacterial infection. In the present study, we cloned and characterized the Tf and TfR genes in Megalobrama amblycephala. The M. amblycephala Tf gene contained 17 exons and 16 introns, encoding 651 amino acids, while the M. amblycephala TfR gene contained 18 exons and 17 introns, encoding 768 amino acids. In healthy fish, Tf mRNA was most abundant in the liver, and TfR was highly expressed in the blood and brain. During early development, the expression of Tf increased from 12 hpf (hour post fertilization) to 26 hpf, followed by a diminution at 32 hpf, then increased significantly to the peak level at 2 dph (day post hatching). The expression pattern of TfR was similar to that of Tf, fluctuating from 0 hpf to 32 hpf and dramatically increasing to the peak at 2 dph. Additionally, both Tf and TfR genes responded to Aeromonas hydrophila infection, by increasing their expression in the liver, spleen and kidney at both mRNA and protein levels, indicating that they were involved in M. amblycephala immune response. Immunohistochemical analysis and Prussian blue staining verified the internalization of TF-receptor system with bound-iron in the liver of M. amblycephala.