Molecular Characterization of E-Type Prostanoid Receptor 4 EP4 from Ayu Plecoglossus altivelis and Its Functional Analysis in the Monocytes-MacrophagesReportar como inadecuado




Molecular Characterization of E-Type Prostanoid Receptor 4 EP4 from Ayu Plecoglossus altivelis and Its Functional Analysis in the Monocytes-Macrophages - Descarga este documento en PDF. Documentación en PDF para descargar gratis. Disponible también para leer online.

Prostaglandin E2 PGE2 plays an important role in a broad spectrum of physiological and pathological processes by interacting with E-type prostanoid receptors EPs. EP4 is one of four EP subtypes known to mediate the immune response in mammalian monocytes-macrophages. However, the precise function and characteristics of EP4 in fish remain unclear. In this study, we characterized a novel EP4-like PaEP4L gene from ayu, Plecoglossus altivelis. The cDNA sequence of PaEP4L is 2781 nucleotides nts in length, encoding a polypeptide of 459 amino acid residues with a calculated molecular weight of 51.17 kDa. Sequence comparison and phylogenetic tree analysis showed that PaEP4L shared 76% amino acid identity with that of the Atlantic salmon Salmo salar. PaEP4L mRNA was detected by real-time quantitative PCR QPCR in all tested tissues and head kidney-derived monocytes-macrophages MO-MФ. It varied greatly in liver, spleen and MO-MФ upon Vibrio anguillarum infection. Western blot analysis revealed a significant increase of PaEP4L in cell homogenates from ayu MO-MФ upon V. anguillarum infection. Moreover, anti-PaEP4L IgG reversed the down-regulation of interleukin 1β IL-1β and tumor necrosis factor α TNF-α mRNA expression as well as phagocytosis in ayu MO-MФ caused by PGE2. There were no significant differences in the respiratory burst response between PGE2 treated and untreated cells. We further found that cAMP mediated PGE2-PaEP4L signal in ayu MO-MФ. In conclusion, our results indicate that PaEP4L mediates PGE2 effects on ayu MO-MФ function, revealing that EP4 also plays a role in the modulation of cells of the fish’s innate immune system.



Autor: Ye-Jing Rong, Xin-Jiang Lu, Jiong Chen

Fuente: http://plos.srce.hr/



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