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AMP-activated protein kinase in the grass carp Ctenopharyngodon idellus: Molecular characterization, tissue distribution and mRNA expression in response to overwinter starvation stress.
Comparative Biochemistry and Physiology B: Biochemistry & Molecular Biology ( IF 1.9 ) Pub Date : 2020-05-15 , DOI: 10.1016/j.cbpb.2020.110457 Wenyi Wu 1 , Jian Sun 1 , Hong Ji 1 , Haibo Yu 1 , Jishu Zhou 1
Comparative Biochemistry and Physiology B: Biochemistry & Molecular Biology ( IF 1.9 ) Pub Date : 2020-05-15 , DOI: 10.1016/j.cbpb.2020.110457 Wenyi Wu 1 , Jian Sun 1 , Hong Ji 1 , Haibo Yu 1 , Jishu Zhou 1
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Adenosine monophosphate-activated protein kinase (AMPK) is the main energy sensor in mammals, but limited information is available regarding its role as an energy sensor in nutrient-restricted fish particularly in period of overwinter starvation. The present study aimed to investigate the role of AMPK in the grass carp Ctenopharyngodon idellus through characterization of AMPK full-length cDNAs and the measurement of transcriptional activity in response to overwinter starvation. AMPK is a heterotrimeric serine/threonine kinase that consists of a catalytic alpha (α) subunit complexed with two regulatory subunits, beta (β) and gamma (γ). In our study, we identified nine isoforms of the AMPK family in grass carp and obtained their complete coding sequences (CDS). In the grass carp, the α subunit is encoded by two isoforms (α1 and α2). The β and γ subunits are encoded by three (β1a, β1b, β2) and four isoforms (γ1, γ2a, γ2b, γ3), respectively. AMPK isoforms in grass carp possess structural features similar to mammalian AMPK and exhibit a high degree of homology with other fish and vertebrate AMPK sequences. The mRNA of nine grass carp AMPK isoforms were found to be expressed in a wide range of tissues in vivo, but the abundance of each AMPK mRNA demonstrated a tissue-dependent expression pattern, indicating that they might be key complexes playing the role of energy metabolism sensors during overwinter starvation conditions. Compared to expression levels in control fish (week 0), the expression of various AMPK isoforms significantly increased in the hepatopancreas of fish exposed to 1 week or more of overwinter starvation conditions as follows: week 1 (AMPK α1 and AMPK α2), week8 (AMPK β1b and AMPK γ2b), week 12 (AMPK β2 and AMPK γ1), and week 16 (AMPK β1a, AMPK γ2a, and AMPK γ3). Additionally, compared to expression levels in control fish (week 0), the expression of various AMPK isoforms significantly increased in the adipose tissue of fish exposed to 1 week or more of overwinter starvation conditions as follows: week 1(AMPK β1a and AMPK β1b), week 4 (AMPK α1, AMPK α2, AMPK γ1, AMPK γ2b and AMPK γ3), and week 8 (AMPK β2 and AMPK γ2a). Further in vitro analysis revealed that the mRNA levels of AMPK isoforms in hepatocytes (AMPK α1, AMPK α2, AMPK β1a, AMPK β1b, AMPK β2, AMPK γ2b and AMPK γ3) and adipocytes (AMPK γ2a, AMPK γ2b and AMPK γ3) changed significantly with in the first 24 h of exposure to the overwinter starvation conditions. These findings confirm that nine AMPK subunits are present in grass carp and that all encode proteins with conserved functional domains. The nine AMPK subunits are all regulated at the transcriptional levels to manage excess energy expenditure during overwinter starvation stress.
中文翻译：
草鱼Ctenopharyngodon idellus中的AMP激活蛋白激酶：响应越冬饥饿压力的分子表征，组织分布和mRNA表达。
腺苷一磷酸激活的蛋白激酶（AMPK）是哺乳动物中的主要能量传感器，但是关于其作为营养受限型鱼中能量传感器的作用的信息很少，特别是在越冬饥饿期。本研究旨在通过表征AMPK全长cDNAs和测量响应越冬饥饿的转录活性来研究AMPK在草鱼Ctenopharyngodon idellus中的作用。AMPK是一种异三聚体丝氨酸/苏氨酸激酶，由与两个调节亚基（β（β）和γ（γ））复合的催化性α（α）亚基组成。在我们的研究中，我们在草鱼中鉴定了AMPK家族的9种亚型，并获得了其完整的编码序列（CDS）。在草鱼中，α亚基由两个亚型（α1和α2）编码。β和γ亚基分别由三个（β1a，β1b，β2）和四个同工型（γ1，γ2a，γ2b，γ3）编码。草鱼中的AMPK亚型具有类似于哺乳动物AMPK的结构特征，并且与其他鱼类和脊椎动物AMPK序列具有高度同源性。发现9种草鱼AMPK亚型的mRNA在体内广泛的组织中表达，但每种AMPK mRNA的丰度均显示出组织依赖性的表达模式，表明它们可能是发挥能量代谢作用的关键复合物越冬饥饿状况下的传感器。与对照鱼（第0周）的表达水平相比，暴露于1周或更长时间的越冬饥饿状况的鱼的肝胰腺中各种AMPK亚型的表达显着增加，如下所示：第1周（AMPKα1和AMPKα2），第8周（AMPKβ1b和AMPKγ2b），第12周（AMPKβ2和AMPKγ1）和第16周（AMPKβ1a，AMPKγ2a和AMPKγ3）。此外，与对照鱼（第0周）的表达水平相比，暴露于1周或更长时间的越冬饥饿状况的鱼的脂肪组织中各种AMPK亚型的表达显着增加，如下所示：第1周（AMPKβ1a和AMPKβ1b） ，第4周（AMPKα1，AMPKα2，AMPKγ1，AMPKγ2b和AMPKγ3）和第8周（AMPKβ2和AMPKγ2a）。进一步的体外分析表明，肝细胞（AMPKα1，AMPKα2，AMPKβ1a，AMPKβ1b，AMPKβ2，AMPKγ2b和AMPKγ3）和脂肪细胞（AMPKγ2a，AMPKγ2b和AMPKγ3）的mRNA水平发生了显着变化。在暴露于冬季饥饿条件的最初24小时内。这些发现证实草鱼中存在9个AMPK亚基，并且全部编码具有保守功能域的蛋白质。九个AMPK亚基均在转录水平受到调控，以控制越冬饥饿压力下的过多能量消耗。
更新日期：2020-05-15
中文翻译：
草鱼Ctenopharyngodon idellus中的AMP激活蛋白激酶：响应越冬饥饿压力的分子表征，组织分布和mRNA表达。
腺苷一磷酸激活的蛋白激酶（AMPK）是哺乳动物中的主要能量传感器，但是关于其作为营养受限型鱼中能量传感器的作用的信息很少，特别是在越冬饥饿期。本研究旨在通过表征AMPK全长cDNAs和测量响应越冬饥饿的转录活性来研究AMPK在草鱼Ctenopharyngodon idellus中的作用。AMPK是一种异三聚体丝氨酸/苏氨酸激酶，由与两个调节亚基（β（β）和γ（γ））复合的催化性α（α）亚基组成。在我们的研究中，我们在草鱼中鉴定了AMPK家族的9种亚型，并获得了其完整的编码序列（CDS）。在草鱼中，α亚基由两个亚型（α1和α2）编码。β和γ亚基分别由三个（β1a，β1b，β2）和四个同工型（γ1，γ2a，γ2b，γ3）编码。草鱼中的AMPK亚型具有类似于哺乳动物AMPK的结构特征，并且与其他鱼类和脊椎动物AMPK序列具有高度同源性。发现9种草鱼AMPK亚型的mRNA在体内广泛的组织中表达，但每种AMPK mRNA的丰度均显示出组织依赖性的表达模式，表明它们可能是发挥能量代谢作用的关键复合物越冬饥饿状况下的传感器。与对照鱼（第0周）的表达水平相比，暴露于1周或更长时间的越冬饥饿状况的鱼的肝胰腺中各种AMPK亚型的表达显着增加，如下所示：第1周（AMPKα1和AMPKα2），第8周（AMPKβ1b和AMPKγ2b），第12周（AMPKβ2和AMPKγ1）和第16周（AMPKβ1a，AMPKγ2a和AMPKγ3）。此外，与对照鱼（第0周）的表达水平相比，暴露于1周或更长时间的越冬饥饿状况的鱼的脂肪组织中各种AMPK亚型的表达显着增加，如下所示：第1周（AMPKβ1a和AMPKβ1b） ，第4周（AMPKα1，AMPKα2，AMPKγ1，AMPKγ2b和AMPKγ3）和第8周（AMPKβ2和AMPKγ2a）。进一步的体外分析表明，肝细胞（AMPKα1，AMPKα2，AMPKβ1a，AMPKβ1b，AMPKβ2，AMPKγ2b和AMPKγ3）和脂肪细胞（AMPKγ2a，AMPKγ2b和AMPKγ3）的mRNA水平发生了显着变化。在暴露于冬季饥饿条件的最初24小时内。这些发现证实草鱼中存在9个AMPK亚基，并且全部编码具有保守功能域的蛋白质。九个AMPK亚基均在转录水平受到调控，以控制越冬饥饿压力下的过多能量消耗。
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