Molecular characterization of adenosine monophosphate deaminase 1 and its regulatory mechanism for inosine monophosphate formation in triploid crucian carp

Inosine monophosphate (IMP) is the main flavoring substance in aquatic animal, and adenosine monophosphate deaminase1 ( AMPD1 ) gene is a key gene in IMP formation. At present, the research on the mechanism of AMPD1 regulating IMP formation in aquatic animal is still blank. In this study, in order t...

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Published inFrontiers in physiology Vol. 13; p. 970939
Main Authors Zhou, Yonghua, Zuo, Anli, Li, Yingjie, Zhang, Yu, Yi, Zilin, Zhao, Dafang, Tang, Jianzhou, Qu, Fufa, Cao, Shenping, Mao, Zhuangwen, Jin, Junyan, Liu, Zhen
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Abstract Inosine monophosphate (IMP) is the main flavoring substance in aquatic animal, and adenosine monophosphate deaminase1 ( AMPD1 ) gene is a key gene in IMP formation. At present, the research on the mechanism of AMPD1 regulating IMP formation in aquatic animal is still blank. In this study, in order to study the mechanism of AMPD1 regulating IMP formation in fish, the full open reading frame (ORF) of AMPD1 which was 2160bp was obtained for the first time in triploid crucian carp ( Carassius auratus ). It encoded 719 amino acids with a molecular mass of 82.97 kDa, and the theoretical isoelectric point value was 6.31. The homology analysis showed that the homology of triploid crucian carp and diploid Carassius auratus w as the highest, up to 99%. And the phylogenetic tree showed that triploid crucian carp was grouped with diploid Carassius auratus , Culter alburnus , and Danio rerio . And real-time fluorescence quantitative results showed that AMPD1 was expressed specifically in muscle of triploid crucian carp ( p < 0.05). The results of detection the localization of AMPD1 in cells indicated that the AMPD1 was mainly localized in cytoplasm and cell membrane. Further, we examined the effects of glutamate which was the promotor of IMP formation on the expression of AMPD1 and the formation of IMP in vivo and in vitro experiments, the results showed that 3% glutamate and 2 mg/ml glutamate could significantly promote AMPD1 expression and IMP formation in triploid crucian carp muscle tissue and muscle cells ( p < 0.05). Then we inhibited the expression of AMPD1 in vivo and in vitro experiments, we found the formation of IMP in muscle tissue and muscle cells of triploid crucian carp all were inhibited and they affected the gene expression of AMPK-mTOR signaling pathway. The all results showed that AMPD1 mediated glutamate through AMPK-mTOR signaling pathway to regulate the formation of fish IMP.
AbstractList Inosine monophosphate (IMP) is the main flavoring substance in aquatic animal, and adenosine monophosphate deaminase1 ( AMPD1 ) gene is a key gene in IMP formation. At present, the research on the mechanism of AMPD1 regulating IMP formation in aquatic animal is still blank. In this study, in order to study the mechanism of AMPD1 regulating IMP formation in fish, the full open reading frame (ORF) of AMPD1 which was 2160bp was obtained for the first time in triploid crucian carp ( Carassius auratus ). It encoded 719 amino acids with a molecular mass of 82.97 kDa, and the theoretical isoelectric point value was 6.31. The homology analysis showed that the homology of triploid crucian carp and diploid Carassius auratus w as the highest, up to 99%. And the phylogenetic tree showed that triploid crucian carp was grouped with diploid Carassius auratus , Culter alburnus , and Danio rerio . And real-time fluorescence quantitative results showed that AMPD1 was expressed specifically in muscle of triploid crucian carp ( p < 0.05). The results of detection the localization of AMPD1 in cells indicated that the AMPD1 was mainly localized in cytoplasm and cell membrane. Further, we examined the effects of glutamate which was the promotor of IMP formation on the expression of AMPD1 and the formation of IMP in vivo and in vitro experiments, the results showed that 3% glutamate and 2 mg/ml glutamate could significantly promote AMPD1 expression and IMP formation in triploid crucian carp muscle tissue and muscle cells ( p < 0.05). Then we inhibited the expression of AMPD1 in vivo and in vitro experiments, we found the formation of IMP in muscle tissue and muscle cells of triploid crucian carp all were inhibited and they affected the gene expression of AMPK-mTOR signaling pathway. The all results showed that AMPD1 mediated glutamate through AMPK-mTOR signaling pathway to regulate the formation of fish IMP.
Inosine monophosphate (IMP) is the main flavoring substance in aquatic animal, and adenosine monophosphate deaminase1 (AMPD1) gene is a key gene in IMP formation. At present, the research on the mechanism of AMPD1 regulating IMP formation in aquatic animal is still blank. In this study, in order to study the mechanism of AMPD1 regulating IMP formation in fish, the full open reading frame (ORF) of AMPD1 which was 2160bp was obtained for the first time in triploid crucian carp (Carassius auratus). It encoded 719 amino acids with a molecular mass of 82.97 kDa, and the theoretical isoelectric point value was 6.31. The homology analysis showed that the homology of triploid crucian carp and diploid Carassius auratus was the highest, up to 99%. And the phylogenetic tree showed that triploid crucian carp was grouped with diploid Carassius auratus, Culter alburnus, and Danio rerio. And real-time fluorescence quantitative results showed that AMPD1 was expressed specifically in muscle of triploid crucian carp (p < 0.05). The results of detection the localization of AMPD1 in cells indicated that the AMPD1 was mainly localized in cytoplasm and cell membrane. Further, we examined the effects of glutamate which was the promotor of IMP formation on the expression of AMPD1 and the formation of IMP in vivo and in vitro experiments, the results showed that 3% glutamate and 2 mg/ml glutamate could significantly promote AMPD1 expression and IMP formation in triploid crucian carp muscle tissue and muscle cells (p < 0.05). Then we inhibited the expression of AMPD1 in vivo and in vitro experiments, we found the formation of IMP in muscle tissue and muscle cells of triploid crucian carp all were inhibited and they affected the gene expression of AMPK-mTOR signaling pathway. The all results showed that AMPD1 mediated glutamate through AMPK-mTOR signaling pathway to regulate the formation of fish IMP.
Author Qu, Fufa
Zuo, Anli
Cao, Shenping
Zhang, Yu
Yi, Zilin
Zhao, Dafang
Mao, Zhuangwen
Liu, Zhen
Zhou, Yonghua
Tang, Jianzhou
Li, Yingjie
Jin, Junyan
AuthorAffiliation 2 State Key Laboratory of Freshwater Ecology and Biotechnology , Institute of Hydrobiology , Chinese Academy of Sciences , Wuhan , China
1 Hunan Provincial Key Laboratory of Nutrition and Quality Control of Aquatic Animals , Department of Biological and Environmental Engineering , Changsha University , Changsha , China
AuthorAffiliation_xml – name: 2 State Key Laboratory of Freshwater Ecology and Biotechnology , Institute of Hydrobiology , Chinese Academy of Sciences , Wuhan , China
– name: 1 Hunan Provincial Key Laboratory of Nutrition and Quality Control of Aquatic Animals , Department of Biological and Environmental Engineering , Changsha University , Changsha , China
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CitedBy_id crossref_primary_10_1007_s10126_023_10270_z
crossref_primary_10_1016_j_cbpb_2024_110966
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This article was submitted to Aquatic Physiology, a section of the journal Frontiers in Physiology
Edited by: Menghong Hu, Shanghai Ocean University, China
Reviewed by: Shuming Zou, Shanghai Ocean University, China
Qizhi Liu, Cedars Sinai Medical Center, United States
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Snippet Inosine monophosphate (IMP) is the main flavoring substance in aquatic animal, and adenosine monophosphate deaminase1 ( AMPD1 ) gene is a key gene in IMP...
Inosine monophosphate (IMP) is the main flavoring substance in aquatic animal, and adenosine monophosphate deaminase1 (AMPD1) gene is a key gene in IMP...
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SubjectTerms AMPD1
glutamate
inosine monophosphate
Physiology
regulatory
triploid crucian carp
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Title Molecular characterization of adenosine monophosphate deaminase 1 and its regulatory mechanism for inosine monophosphate formation in triploid crucian carp
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https://pubmed.ncbi.nlm.nih.gov/PMC9468423
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Volume 13
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