AUTHOR=Zhou Yonghua , Zuo Anli , Li Yingjie , Zhang Yu , Yi Zilin , Zhao Dafang , Tang Jianzhou , Qu Fufa , Cao Shenping , Mao Zhuangwen , Jin Junyan , Liu Zhen 

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

JOURNAL=Frontiers in Physiology

VOLUME=13

YEAR=2022

URL=https://www.frontiersin.org/journals/physiology/articles/10.3389/fphys.2022.970939

DOI=10.3389/fphys.2022.970939

ISSN=1664-042X

ABSTRACT=<p>Inosine monophosphate (IMP) is the main flavoring substance in aquatic animal, and adenosine monophosphate deaminase1 (<italic>AMPD1</italic>) gene is a key gene in IMP formation. At present, the research on the mechanism of <italic>AMPD1</italic> regulating IMP formation in aquatic animal is still blank. In this study, in order to study the mechanism of <italic>AMPD1</italic> regulating IMP formation in fish, the full open reading frame (ORF) of <italic>AMPD1</italic> which was 2160bp was obtained for the first time in triploid crucian carp (<italic>Carassius auratus</italic>). 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 <italic>Carassius auratus w</italic>as the highest, up to 99%. And the phylogenetic tree showed that triploid crucian carp was grouped with diploid <italic>Carassius auratus</italic>, <italic>Culter alburnus</italic>, and <italic>Danio rerio</italic>. And real-time fluorescence quantitative results showed that <italic>AMPD1</italic> was expressed specifically in muscle of triploid crucian carp (<italic>p</italic> &lt; 0.05). The results of detection the localization of <italic>AMPD1</italic> in cells indicated that the <italic>AMPD1</italic> 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 <italic>AMPD1</italic> and the formation of IMP <italic>in vivo</italic> and <italic>in vitro</italic> experiments, the results showed that 3% glutamate and 2 mg/ml glutamate could significantly promote <italic>AMPD1</italic> expression and IMP formation in triploid crucian carp muscle tissue and muscle cells (<italic>p</italic> &lt; 0.05). Then we inhibited the expression of <italic>AMPD1 in vivo</italic> and <italic>in vitro</italic> 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 <italic>AMPD1</italic> mediated glutamate through AMPK-mTOR signaling pathway to regulate the formation of fish IMP.</p>