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Molecular Characterization of E-Type Prostanoid Receptor 4 (EP4) from Ayu (Plecoglossus altivelis) and Its Functional Analysis in the Monocytes/Macrophages
- Source :
- PLoS ONE, Vol 11, Iss 1, p e0147884 (2016), PLoS ONE
- Publication Year :
- 2016
- Publisher :
- Public Library of Science (PLoS), 2016.
-
Abstract
- 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.
- Subjects :
- 0301 basic medicine
Vibrio anguillarum
Physiology
lcsh:Medicine
Biochemistry
Monocytes
White Blood Cells
0302 clinical medicine
Animal Cells
Immune Physiology
Medicine and Health Sciences
Receptor
lcsh:Science
Immune Response
Zebrafish
Respiratory Burst
Innate Immune System
Multidisciplinary
medicine.diagnostic_test
Fishes
Animal Models
Complementary DNA
Respiratory burst
Nucleic acids
medicine.anatomical_structure
Cell Processes
Osteichthyes
Osmeriformes
030220 oncology & carcinogenesis
Vertebrates
Cytokines
lipids (amino acids, peptides, and proteins)
Cellular Types
Research Article
Multiple Alignment Calculation
Forms of DNA
Immune Cells
Immunology
Spleen
Biology
Research and Analysis Methods
03 medical and health sciences
Model Organisms
Extraction techniques
Immune system
Phagocytosis
Western blot
Computational Techniques
Genetics
medicine
Animals
Blood Cells
Innate immune system
Macrophages
lcsh:R
Organisms
Biology and Life Sciences
Cell Biology
DNA
Molecular Development
biology.organism_classification
Molecular biology
Split-Decomposition Method
RNA extraction
030104 developmental biology
Immune System
lcsh:Q
Plecoglossus altivelis
Receptors, Prostaglandin E, EP4 Subtype
Developmental Biology
Subjects
Details
- Language :
- English
- ISSN :
- 19326203
- Volume :
- 11
- Issue :
- 1
- Database :
- OpenAIRE
- Journal :
- PLoS ONE
- Accession number :
- edsair.doi.dedup.....fea4735699bdea77b9e6bac6a4a23da9