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The immunomodulation mediated by a delta-opioid receptor for [Met5]-enkephalin in oyster Crassostrea gigas

Liu, Zhaoqun, Zhou, Zhi, Wang, Lingling, Jiang, Shuai, Wang, Weilin, Zhang, Ran, Song, Linsheng
Developmental and comparative immunology 2015 v.49 no.2 pp. 217-224
Crassostrea gigas, G-protein coupled receptors, amino acids, antagonists, antibacterial properties, calcium, complementary DNA, cyclic AMP, gills, hemocytes, hepatopancreas, homeostasis, immune response, immunocytes, immunomodulation, kidneys, ligands, messenger RNA, muscles, narcotics, oysters, pathogens, second messengers
Opioid receptors (OR) are a group of G protein-coupled receptors with opioids as ligands, which play an important role in triggering the second messengers to modulate immune response in vertebrate immunocytes. In the present study, the full length cDNA of a homologue of δ-opioid receptor (DOR) for [Met5]-enkaphalin was cloned from oyster Crassostrea gigas (designated as CgDOR), which was 1104 bp encoding a peptide of 367 amino acids containing a conserved 7tm_1 domain. After the stimulation of [Met5]-enkephalin, the concentration of second messengers Ca2+ and cAMP in the HEK293T cells decreased significantly (p < 0.05) with the expression of CgDOR. However, this trend was reverted with the addition of DOR antagonist BNTX. The CgDOR transcripts were ubiquitously detected in the tested tissues including haemocytes, gonad, mantle, kidney, gill, adductor muscle and hepatopancreas, with the highest expression level in the hepatopancreas. After LPS stimulation, the expression level of CgDOR mRNA began to increase (4.05-fold, p < 0.05) at 6 h, and reached the highest level (5.00-fold, p < 0.05) at 12 h. Haemocyte phagocytic and antibacterial activities increased significantly after [Met5]-enkephalin stimulation, whereas the increase was repressed with the addition of DOR antagonist BNTX. These results collectively suggested that CgDOR for [Met5]-enkephalin could modulate the haemocyte phagocytic and antibacterial functions through the second messengers Ca2+ and cAMP, which might be requisite for pathogen elimination and homeostasis maintenance in oyster.