Molecular cloning, characterization and effects of catechol-omethyltransferase (comt) mrna and protein on aggressive behavior in the swimming crab portunus trituberculatus
Molecular cloning, characterization and effects of catechol-omethyltransferase (comt) mrna and protein on aggressive behavior in the swimming crab portunus trituberculatus
复制标题
儿茶酚甲基转移酶 (comt) mRNA 和蛋白的分子克隆、表征及其对三疣梭子蟹攻击行为的影响
DOI：
10.1016/j.aquaculture.2018.06.055
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发表时间：
2018
期刊：
影响因子：
4.5
通讯作者：
Wang Chunlin
中科院分区：
文献类型：
--
作者：
Zhu Fang;Fu Yuanyuan;Mu Changkao;Liu Lei;Li Ronghua;Song Weiwei;Shi Ce;Ye Yangfang;Wang Chunlin
本研究从三疣梭子蟹（Portunus trituberculatus）肌肉中克隆了一个编码儿茶酚-O-甲基转移酶（catechol-O-methyltransferase，COMT）的cDNA，命名为PtCOMT，PtCOMT是一个保守的基因，编码一个由222个氨基酸组成的蛋白质，分子量为25.4 kDa，仅由“SAM_OMT_I”组成。RT-PCR结果表明，PtCOMT基因在蚤状幼体Ⅰ期（Z1）、蚤状幼体Ⅱ期（Z2）、蚤状幼体Ⅲ期（Z3）、蚤状幼体Ⅳ期（Z4）和大眼幼体（M）中均有表达。三疣虫。PtCOMTmRNA表达在Z4期和M期高于Z1、Z2和Z3期（P< 0.05）。此外，PtCOMT在健康蟹的组织中表达，即神经节、肌肉和肝胰腺。RT-PCR和免疫组化结果显示，在神经节和肌肉中的表达量显著高于肝胰腺（P< 0.05）。DA注射蟹的侵略性进行了测试，通过行为研究，多巴胺处理的螃蟹走通过“中间”的池比对照螃蟹更频繁。与对照组相比，注射多巴胺（DA）后2 h，肌肉中PtCOMT表达显著上调（P< 0.05），4 h后表达显著下调（P< 0.05）;注射多巴胺后2 h，神经节中PtCOMT表达显著下调（P<0.05），4 h后表达显著上调（P< 0.05）。这些结果表明DA促进PtCOMT的释放。我们的研究结果表明PtCOMT的表达水平可能与攻击性直接相关，并影响中枢神经系统。trituberculatusto行动on its aggressive侵略behavior行为.本研究结果为进一步研究白蛉的攻击行为机制提供了理论依据。三疣虫
In the present study, a cDNA that encodes catechol-O-methyltransferase (COMT) was cloned from the muscle of the swimming crabPortunus trituberculatusand namedPtCOMT.PtCOMTwas proven to be a conserved gene that encodes a protein of 222 amino acid residues with a molecular mass 25.4 kDa that constitutes only the signature motif “SAM_OMT_I.” The RT-PCR results showed thatPtCOMTwas expressed in all five larval stages (zoea stage I, Z1; zoea stage II, Z2; zoea stage III, Z3; zoea stage IV, Z4; and megalopa, M) ofP. trituberculatus. ThePtCOMTmRNA levels were higher (P< 0.05) in the Z4and M stages than in the Z1, Z2, and Z3stages. In addition,PtCOMTwas expressed in the tissues of healthy crabs, namely, ganglion, muscle, and hepatopancreas. On the basis of the RT-PCR and immunohistochemistry analyses, the level of expression in the ganglion and muscle was significantly higher than that in the hepatopancreas (P< 0.05). Aggressiveness of DA injected crabs was tested through a behavioral study where dopamine-treated crabs walked through the “middle” of the pool more frequently than control crabs. When compared with the control group,PtCOMTexpression level was significantly upregulated (P< 0.05) at 2 h and significantly downregulated at 4 h in the muscles after dopamine (DA) injection (P< 0.05), whereas it was significantly downregulated (P< 0.05) at 2 h and upregulated after 4 h (P< 0.05) in the ganglia. These results show that DA promotes the release ofPtCOMT. Our findings indicate that the expression level ofPtCOMTmay be directly related to aggressiveness and affect the central nervous system ofP. trituberculatusto act on its aggressive behavior. Our results can help further studies on the aggressive behavior mechanism ofP. trituberculatus.