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An amine oxidase gene from mud crab, Scylla paramamosain, regulates the neurotransmitters serotonin and dopamine in vitro.

ABSTRACT: Amine oxidase, which participates in the metabolic processing of biogenic amines, is widely found in organisms, including higher organisms and various microorganisms. In this study, the full-length cDNA of a novel amine oxidase gene was cloned from the mud crab, Scylla paramamosain, and termed SpAMO. The cDNA sequence was 2,599 bp in length, including an open reading frame of 1,521 bp encoding 506 amino acids. Two amino acid sequence motifs, a flavin adenine dinucleotide-binding domain and a flavin-containing amine oxidoreductase, were highly conserved in SpAMO. A quantitative real-time polymerase chain reaction analysis showed that the expression level of SpAMO after quercetin treatment was time- and concentration-dependent. The expression of SpAMO tended to decrease and then increase in the brain and haemolymph after treatment with 5 mg/kg/d quercetin; after treatment with 50 mg/kg/d quercetin, the expression of SpAMO declined rapidly and remained low in the brain and haemolymph. These results indicated that quercetin could inhibit the transcription of SpAMO, and the high dose (50 mg/kg/d) had a relatively significant inhibitory effect. SpAMO showed the highest catalytic activity on serotonin, followed by dopamine, ?-phenylethylamine, and spermine, suggesting that the specific substrates of SpAMO are serotonin and dopamine. A bioinformatics analysis of SpAMO showed that it has molecular characteristics of spermine oxidase, but a quercetin test and enzyme activity study indicated that it also functions like monoamine oxidase. It is speculated that SpAMO might be a novel amine oxidase in S. paramamosain that has the functions of both spermine oxidase and monoamine oxidase.

SUBMITTER: Liu J

PROVIDER: S-EPMC6152983 | biostudies-literature | 2018

REPOSITORIES: biostudies-literature

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An amine oxidase gene from mud crab, Scylla paramamosain, regulates the neurotransmitters serotonin and dopamine in vitro.

Liu Junguo J Zhao Ming M Song Wei W Ma Lingbo L Li Xiu X Zhang Fengying F Diao Le L Pi Yan Y Jiang Keji K

PloS one 20180924 9

Amine oxidase, which participates in the metabolic processing of biogenic amines, is widely found in organisms, including higher organisms and various microorganisms. In this study, the full-length cDNA of a novel amine oxidase gene was cloned from the mud crab, Scylla paramamosain, and termed SpAMO. The cDNA sequence was 2,599 bp in length, including an open reading frame of 1,521 bp encoding 506 amino acids. Two amino acid sequence motifs, a flavin adenine dinucleotide-binding domain and a fla ...[more]

PMID: 30248122

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Project description:BackgroundInfection of bacterial Vibrio parahaemolyticus is common in mud crab farms. However, the mechanisms of the crab's response to pathogenic V. parahaemolyticus infection are not fully understood. MicroRNAs (miRNAs) are a class of small noncoding RNAs that function as regulators of gene expression and play essential roles in various biological processes. To understand the underlying mechanisms of the molecular immune response of the crab to the pathogens, high-throughput Illumina/Solexa deep sequencing technology was used to investigate the expression profiles of miRNAs in S. paramamosain under V. parahaemolyticus infection.Methodology/principal findingsTwo mixed RNA pools of 7 tissues (intestine, heart, liver, gill, brain, muscle and blood) were obtained from V. parahaemolyticus infected crabs and the control groups, respectively. By aligning the sequencing data with known miRNAs, we characterized 421 miRNA families, and 133 conserved miRNA families in mud crab S. paramamosain were either identical or very similar to existing miRNAs in miRBase. Stem-loop qRT-PCRs were used to scan the expression levels of four randomly chosen differentially expressed miRNAs and tissue distribution. Eight novel potential miRNAs were confirmed by qRT-PCR analysis and the precursors of these novel miRNAs were verified by PCR amplification, cloning and sequencing in S. paramamosain. 161 miRNAs (106 of which up-regulated and 55 down-regulated) were significantly differentially expressed during the challenge and the potential targets of these differentially expressed miRNAs were predicted. Furthermore, we demonstrated evolutionary conservation of mud crab miRNAs in the animal evolution process.Conclusions/significanceIn this study, a large number of miRNAs were identified in S. paramamosain when challenged with V. parahaemolyticus, some of which were differentially expressed. The results show that miRNAs might play some important roles in regulating gene expression in mud crab under V. parahaemolyticus infection, providing a basis for further investigation of miRNA-modulating networks in innate immunity of mud crab.

Project description:Viruses associated with sleeping disease (SD) in crabs cause great economic losses to aquaculture, and no effective measures are available for their prevention. In this study, to help develop novel antiviral strategies, single-particle cryo-electron microscopy was applied to investigate viruses associated with SD. The results not only revealed the structure of mud crab dicistrovirus (MCDV) but also identified a novel mud crab tombus-like virus (MCTV) not previously detected using molecular biology methods. The structure of MCDV at a 3.5-Å resolution reveals three major capsid proteins (VP1 to VP3) organized into a pseudo-T=3 icosahedral capsid, and affirms the existence of VP4. Unusually, MCDV VP3 contains a long C-terminal region and forms a novel protrusion that has not been observed in other dicistrovirus. Our results also reveal that MCDV can release its genome via conformation changes of the protrusions when viral mixtures are heated. The structure of MCTV at a 3.3-Å resolution reveals a T= 3 icosahedral capsid with common features of both tombusviruses and nodaviruses. Furthermore, MCTV has a novel hydrophobic tunnel beneath the 5-fold vertex and 30 dimeric protrusions composed of the P-domains of the capsid protein at the 2-fold axes that are exposed on the virion surface. The structural features of MCTV are consistent with a novel type of virus.IMPORTANCE Pathogen identification is vital for unknown infectious outbreaks, especially for dual or multiple infections. Sleeping disease (SD) in crabs causes great economic losses to aquaculture worldwide. Here we report the discovery and identification of a novel virus in mud crabs with multiple infections that was not previously detected by molecular, immune, or traditional electron microscopy (EM) methods. High-resolution structures of pathogenic viruses are essential for a molecular understanding and developing new disease prevention methods. The three-dimensional (3D) structure of the mud crab tombus-like virus (MCTV) and mud crab dicistrovirus (MCDV) determined in this study could assist the development of antiviral inhibitors. The identification of a novel virus in multiple infections previously missed using other methods demonstrates the usefulness of this strategy for investigating multiple infectious outbreaks, even in humans and other animals.

Dataset Information

An amine oxidase gene from mud crab, Scylla paramamosain, regulates the neurotransmitters serotonin and dopamine in vitro.

Publications

An amine oxidase gene from mud crab, Scylla paramamosain, regulates the neurotransmitters serotonin and dopamine in vitro.

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