Expression characteristics and regulatory mechanism of Apela gene in liver of chicken (Gallus gallus)

Apela, a novel endogenous peptide ligand for the G-protein-coupled apelin receptor, was first discovered and identified in human embryonic stem cells in 2013. Apela has showed some biological functions in promoting angiogenesis and inducing vasodilatation of mammals by binding apelin receptor, but little is known about its expression characteristics and regulatory mechanism in chicken. In the present study, the coding sequences of Apela in chicken was cloned. The evolution history and potential function of Apela were analyzed. Subsequently, the spatiotemporal expression characteristics of chicken Apela were investigated. Furthermore, the regulatory mechanism of Apela mRNA responsing to estrogen was explored by in vitro and in vivo experiments. The results showed that the length of the CDs of Apela mRNA was 165 bp and encoded a protein consisting of 54 amino acids residues with a transmembrane domain in chicken. The Apela was derived from the same ancestor of Apelin, and abundantly expressed in liver, kidney and pancreas tissues. The expression levels of Apela in the liver of hens were significantly higher at the peak-laying stage than that at the pre-laying stage (p ≤ 0.05). The Apela mRNA levels were significantly up-regulated in primary hepatocytes treated with 17β-estradiol (p ≤ 0.05), and could be effectively inhibited by estrogen receptor antagonists MPP, ICI 182780 and tamoxifen. It indicated that chicken Apela expression was regulated by estrogen via estrogen receptor α (ERα). In individual levels, both the contents of TG, TC and VLDL-c in serum, and the expression of ApoVLDLII and Apela in liver markedly up-regulated by 17β-estradiol induction at 1mg/kg and 2mg/kg concentrations (p ≤ 0.05). This study lays a foundation for further research on Apela involving in hepatic lipid metabolism.