Cloning and functional characterization of the DA2 receptor gene in Chinese mitten crab (Eriocheir sinensis)

Xiaozhen Yang; Genyong Huang; Min-Jie Xu; Cong Zhang; Yongxu Cheng; Zhigang Yang

doi:10.1371/journal.pone.0193999

Cloning and functional characterization of the DA2 receptor gene in Chinese mitten crab (Eriocheir sinensis)

PLoS One. 2018 Mar 19;13(3):e0193999. doi: 10.1371/journal.pone.0193999. eCollection 2018.

Authors

Xiaozhen Yang^{1

2

3}, Genyong Huang^{1

2

3}, Min-Jie Xu^{1

2

3}, Cong Zhang^{1

2

3}, Yongxu Cheng^{1

2

3}, Zhigang Yang^{1

2

3}

Affiliations

¹ National Demonstration Center for Experimental Fisheries Science Education, Shanghai Ocean University, Shanghai, China.
² Key Laboratory of Freshwater Aquatic Genetic Resources, Ministry of Agriculture, Shanghai Ocean University, Shanghai, China.
³ Shanghai Engineering Research Center of Aquaculture; Shanghai Ocean University, Shanghai, China.

Abstract

Dopamine (DA) plays a modulatory role in numerous physiological processes such as light adaptation and food intake, and exerts these functions through DA receptors (DARs). This study presents, for the first time, isolation and characterization of the dopamine receptor 2(DA2 receptor) cDNA from the intestinal tissue of Eriocheir sinensis, an economically important freshwater aquaculture species in China. The DA2 receptor cDNA sequence, which was obtained by rapid amplification of cDNA ends, is 2369bp long, encode peptide of 589 amino acid, and is highly homologous to related sequences in crustaceans. Analysis of the deduced amino acid sequence and the structure of the DA2 indicated that this receptor is a member of the family of G protein-coupled receptors (GPCRs), as it contains seven transmembrane domains and other common signatures of GPCRs. RT-PCR showed that the expression of the DA2 receptor gene was distributed in various tissues, and high expression levels were observed in the cranial ganglia and the thoracic ganglia. Further study of the effect of photoperiod on DA2 expression showed that constant darkness induced a significant increase in DA2 expression in the cranial ganglia. Finally, analysis of DA2 receptor expression under different feeding statuses showed that there was significantly greater expression in the hepatopancreas and intestines after feeding than before feeding, but there were no differences in expression between the before feeding and during feeding periods in either tissue. Our results indicate that the DA2 receptor structurally belongs to the family of G protein-coupled receptors, and that the cranial ganglia are the main tissues in which the DA2 receptor participates in light adaptation during dark hours. In addition, the DA2 receptor in E. sinensis may be involved in the physiological regulation of the hepatopancreas and digestive tract after the ingestion of food. This study provides a foundation for further exploration of the light adaptation and digestive functions of the DA2 receptor in decapods.

Publication types

Research Support, N.I.H., Extramural

MeSH terms

Amino Acid Sequence
Animals
Base Sequence
Brachyura / genetics*
China
Cloning, Molecular / methods
DNA, Complementary / genetics
Gene Expression Profiling / methods
Gene Expression Regulation / genetics
Hepatopancreas / metabolism
Intestinal Mucosa / metabolism
Phylogeny
Receptors, Dopamine D2 / genetics*
Receptors, G-Protein-Coupled / genetics
Sequence Alignment

Substances

DNA, Complementary
Receptors, Dopamine D2
Receptors, G-Protein-Coupled

Grants and funding

This work was supported by the National Natural Science Foundation of China [grant numbers 31272677, 31472287], the Extension of Chinese Mitten Crab Eriocheir Sinensis Aquaculture Technology Fund of Shanghai Agricultural Commission [grant number 2015D1-7], the Aquaculture Engineering Research Platform in Shanghai Established by Shanghai Science and Technology Commission [grant number 16DZ2281200]), and the China Agriculture Research System-47.