The cholinergic immune regulation mediated by a novel muscarinic acetylcholine receptor through TNF pathway in oyster Crassostrea gigas

Muscarinic receptors, which selectively take muscarine as their ligand, are critical for the immunological and physiological processes in animals. In the present study, the open region frame (ORF) of a homologue of muscarinic acetylcholine (ACh) receptor (mAChR) was amplified from oyster Crassostrea...

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Bibliographic Details
Published in:Developmental & Comparative Immunology
Main Authors: Liu, Zhaoqun, Zhou, Zhi, Wang, Lingling, Dong, Wenjing, Qiu, Limei, Song, Linsheng
Format: Article in Journal/Newspaper
Language:English
Published: 2016
Subjects:
Crassostrea Gigas
Acetylcholine
Muscarinic Acetylcholine Receptor
Tnf Pathway
Immune Regulation
Crassostrea gigas
Online Access:http://ir.qdio.ac.cn/handle/337002/135892
https://doi.org/10.1016/j.dci.2016.07.003
Description
Summary:Muscarinic receptors, which selectively take muscarine as their ligand, are critical for the immunological and physiological processes in animals. In the present study, the open region frame (ORF) of a homologue of muscarinic acetylcholine (ACh) receptor (mAChR) was amplified from oyster Crassostrea gigas (named as CgmAChR-1), whose full length was 1983 bp and the protein it encoded contained 660 amino acids with a seven transmembrane region. Phylogeny analysis suggested that CgmAChR-1 shared homology with M-5 muscarinic receptor found in invertebrates including Habropoda laboriosa, Acromyrmex echinatior and Echinococcus granulosus. After cell transfection of CgrnAChR-1 into HEK293T cells and ACh incubation, the level of intracellular Ca2+ and CAMP increased significantly (p < 0.05). Such trend could be reverted with the addition of M-3 and M-5 muscarinic receptor antagonists DAMP and DAR. The CgmAChR-1 transcripts were ubiquitously detectable in seven different tissues with the maximal expression level in adductor muscle. When the oysters received LPS stimulation, CgmAChR-1 mRNA expression in haemocyte was increased to the highest level (6.05-fold, p < 0.05) at 24 h, while blocking CgmAChR-1 using receptor antagonists before LPS stimulation promoted the expression of oyster TNF, resulting in the increase of haemocyte apoptosis index. These results suggested that CgmAChR-1 was the key molecule in cholinergic neuroendocrine-immune system contributing to the regulation of TNF expression and apoptosis process. (C) 2016 Elsevier Ltd. All rights reserved.

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