INSECT BIOCHEMISTRY AND MOLECULAR BIOLOGY
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2025
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IF 3.7
Shiyan Liu,Xuefeng Zhang,Xin Gong,Jinxin Yu,Tao Lin,Qian Xiang,Xinnian Zeng,Jiali Liu
Dopamine (DA) is a critical molecule within the insect nervous system, known to regulate a myriad of physiological functions and instigate behavioral shifts in insects. It exerts its effects by interacting with specific dopamine receptors (DARs). In this study, three DARs cDNAs from Bactrocera dorsalis (Hendel) (Diptera: Tephritidae) ( BdDOP1 , BdDOP2 and BdDOP3 ) were cloned using molecular biology techniques. These receptors exhibited high sequence identity with their orthologous DARs, and phylogenetic analyses also clustered these receptors within their respective receptor subtype. Additionally, the high expression levels of these DARs in the head suggest their prominent role in the central nervous system of B. dorsalis . To investigate the pharmacological properties of these receptors, expression vectors for BdDOP1 , BdDOP2 and BdDOP3 were constructed and expressed in HEK-293T cells. Our results demonstrated that DA and synthetic agonists activated these receptors in a dose-dependent manner, and DA activation can be competitively inhibited by various antagonists, exhibiting distinct potencies for each dopamine receptor type. Among the tested antagonists, SCH-23390, methiothepin, and metoclopramide were identified as the most potent inhibitors of BdDOP1 , BdDOP2 and BdDOP3 , respectively. This study provides valuable insights into the molecular and pharmacological characteristics of DARs in B. dorsalis , offering a theoretical foundation for the development of novel behavioral modulators targeting these receptors. The findings also serve as a reference for the functional analyses of DARs in other insect species.
研究领域:
昆虫学
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神经生物学
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分子生物学
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DOI
PubMed
高影响因子
FISH PHYSIOLOGY AND BIOCHEMISTRY
·
2020
·Vol. 46
·期 4
·1207-1218
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IF 2.249
Deng Dapeng,Yan Xiao,Zhao Wenli,Qin Chaobin,Yang Guokun,Nie Guoxing
Glucose transporter 2 ( glut2 ) has been studied in mammals, aves, and several fish, while the comparative studies of glut2 in common carp are still lacking. In this study, glut2 was firstly isolated and characterized from the liver of common carp. The full-length cDNA of glut2 was 2351 bp with an open reading frame (ORF) of 1512 bp, encoding 503 amino acids. Alignment of glut2 amino acid sequences from different species revealed that common carp glut2 showed higher sequence identity with teleosts, and lower homology with mammals and amphibians. Tissue distribution demonstrated that glut2 mRNA level was mainly expressed in liver, foregut, and midgut. To investigate the actions of glut2 on glucose metabolism, the level of glut2 mRNA was detected after intraperitoneal injection of glucose, human insulin and glucagon (100 ng/g), respectively. Following glucose administration, glut2 gene expression was significantly upregulated at 3 h in the foregut. However, no change was found in hepatic glut2 mRNA level, indicating that glut2 may have a role in intestinal glucose uptake rather than in the liver. Following insulin treatment, the expression of glut2 was markedly downregulated at 3 h and 6 h in the liver, and at 3 h in the foregut, respectively. Furthermore, glut2 mRNA expression was unaffected by glucagon injection in the liver and foregut. These results suggested that the expression of glut2 regulated by pancreatic hormones was different. Taken together, our studies firstly revealed the structure of the glut2 gene and its potential functions in glucose metabolism of common carp.
研究领域:
分子生物学
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比较生理学
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水产养殖
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内分泌学
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DOI
PubMed
高影响因子
Genes
·
2022
·Vol. 13
·期 7
·1113
·
IF 4.141
Biying Jiang,Baolong Cao,Zhichun Zhou,Zejiao Li,Can Lv,Jiannan Zhang,Heyuan Zhang,Yajun Wang,Juan Li
Adrenoceptors are suggested to mediate the functions of norepinephrine (NE) and epinephrine (EPI) in the central nervous system (CNS) and peripheral tissues in vertebrates. Compared to mammals, the functionality and expression of adrenoceptors have not been well characterized in birds. Here, we reported the structure, expression, and functionality of chicken functional α2A-adrenoceptor, namedADRA2A. The cloned chickenADRA2AcDNA is 1335 bp in length, encoding the receptor with 444 amino acids (a.a.), which shows high amino acid sequence identity (63.4%) with its corresponding ortholog in humans. Using cell-based luciferase reporter assays and Western blot, we demonstrated that the ADRA2A could be activated by both NE and EPI through multiple signaling pathways, including MAPK/ERK signaling cascade. In addition, the mRNA expression ofADRA2Ais found to be expressed abundantly in adult chicken tissues including thyroid, lung, ovary and adipose from the reported RNA-Seq data sets. Moreover, the mRNA expression ofADRA2Ais also found to be highly expressed in the granulosa cells of 6–8 mm and F5 chicken ovarian follicles, which thus supports that ADRA2A signaling may play a role in ovarian follicular growth and differentiation. Taken together, our data provide the first proof that the α2A-adrenoceptor is functional in birds involving avian ovarian follicular development.Keywords:chickens;α2-adrenoceptor;ADRA2A
研究领域:
分子生物学
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内分泌学
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生殖生物学
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DOI
PubMed
高影响因子
Molecular Catalysis
·
2024
·
IF 3.9
Kaixuan Ke,Ling Guo,Zhipeng Qi,Dou Dou,Han Ma,Xianying Fang,Linguo Zhao
Carotenoid cleavage dioxygenases (CCDs) play a pivotal role in the biosynthesis of volatile apocarotenoids, which have significant industrial applications due to their aromatic and bioactive properties. This study focuses on the molecular cloning and biochemical characterization of the LbCCD1 from Lycium barbarum . The LbCCD1 gene was successfully cloned and heterologously expressed in Escherichia coli and several carotenoids were using as substrates for investigate its specificity by in vitro and in vivo experiment. The LbCCD1 protein was able to cleave a variety of carotenoids including β-carotene, zeaxanthin, astaxanthin, and β-apo-8′-carotenal, at the 9, 10 (9′, 10′) double bond to produce β-ionone, 3‑hydroxy-4-oxo-β-ionone, and 3‑hydroxy-β-ionone, respectively in vitro. LbCCD1 could also cleave zeaxanthin and β-carotene at the 9, 10 (9′, 10′) double bond to produce β-ionone, respectively, in E. coli accumulating carotenoids. Interestingly, LbCCD1 did not exhibit cleavage activity on lycopene either in vivo or in vitro unlike other CCD1 family enzymes. In the previous experiment, it was confirmed that LbCCD1 exhibits cleavage activity towards β-apo-8′-carotenal in vitro, so we used β-apo-8′-carotenal as the substrate for characterizing the enzymatic properties. The expression of LbCCD1 was optimized at such conditions (temperature 24 °C, IPTG 0.1 mM, induction time 24 h). The biochemical characterization of LbCCD1 revealed the optimal activities were at pH 9 and 55 °C. The addition of 10 % ethanol could increase enzyme activity to above 15 %. However, the concentration of Fe 2+ has a minimal effect on enzyme activity. The V max for β-apo-8′-carotenal was 8.6 U/mg, while the K m was 0.27 mM. To preliminarily verify the potential of LbCCD1 as a biological component for β-ionone production. By introducing LbCCD1 into the β-carotene-high-producing chassis cell and optimizing the conditions (temperature 30 °C, IPTG 0.01 mM, Fe 2+ concentration 0.05 mM), the β-ionone yield reached 21.45 mg/L. This study focused on one of the CCDs derived from woody plants, which have been relatively underexplored. It lays the groundwork for expanding the CCD enzyme library, identifying suitable CCDs, and investigating the structure-function relationship of CCDs. Furthermore, it sets the stage for engineering novel CCD genes and developing advanced applications of CCDs as biocatalysts and platforms for synthetic biology. These advancements will enable the efficient production of volatile aroma compounds from carotenoids.
研究领域:
植物生物化学
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分子生物学
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生物技术
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合成生物学
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产品关联
DOI
高影响因子
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