{"title":"暴露于不同盐度的日本对虾丙氨酸氨基转移酶基因的克隆和表达谱。","authors":"Hiroki Koyama, Kyoko Yamashita, Hinano Narita, Haruki Hiraoka, Yuka Sasaki, Kanna Kamiya, Rin Yamakawa, Hisato Kuniyoshi, Sanit Piyapattanakorn, Shugo Watabe","doi":"10.1002/jez.2811","DOIUrl":null,"url":null,"abstract":"<p>Several crustaceans including shrimps change the amount of specific free amino acids to regulate the osmotic pressure in their bodies. Kuruma shrimp <i>Penaeus japonicus</i> also increases the concentration of alanine (Ala) in the abdominal muscle following the increase of environmental salinity. In the present study, to elucidate the mechanisms of changes in Ala accumulation of kuruma shrimp depending on salinity, we cloned the gene encoding alanine aminotransferase (ALT), an enzyme involved in Ala biosynthesis, and examined its expression profile. It was found that the full-length kuruma shrimp ALT1 cDNA consisted of 3,301 bp, encoding 514 amino acids, and that all amino acid residues important for ALT activity were conserved. Phylogenetic analysis also indicated that the ALT gene cloned in this study was classified as ALT1. Moreover, we examined the expression levels of the ALT1 gene in the abdominal muscle and the hepatopancreas of kuruma shrimp acclimated at 17‰, 34‰, and 40‰ salinities, resulting that the mRNA levels of the ALT1 genes in both tissues of the shrimp acclimated at 40‰ were significantly higher than those at 17‰ for 12 h (<i>p</i> < 0.05). The mRNA levels of the ALT1 gene in the abdominal muscle of the shrimp acclimated for more than 24 h tended to increase following the increase of environmental salinity. These results indicate that ALT1 is responsible for the increase of free Ala concentration in the abdominal muscle of kuruma shrimp to regulate osmotic pressure at high salinity.</p>","PeriodicalId":15711,"journal":{"name":"Journal of experimental zoology. Part A, Ecological and integrative physiology","volume":"341 5","pages":"615-626"},"PeriodicalIF":1.9000,"publicationDate":"2024-03-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Cloning and expression profile of the alanine aminotransferase gene from kuruma shrimp Penaeus japonicus exposed to different salinities\",\"authors\":\"Hiroki Koyama, Kyoko Yamashita, Hinano Narita, Haruki Hiraoka, Yuka Sasaki, Kanna Kamiya, Rin Yamakawa, Hisato Kuniyoshi, Sanit Piyapattanakorn, Shugo Watabe\",\"doi\":\"10.1002/jez.2811\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p>Several crustaceans including shrimps change the amount of specific free amino acids to regulate the osmotic pressure in their bodies. Kuruma shrimp <i>Penaeus japonicus</i> also increases the concentration of alanine (Ala) in the abdominal muscle following the increase of environmental salinity. In the present study, to elucidate the mechanisms of changes in Ala accumulation of kuruma shrimp depending on salinity, we cloned the gene encoding alanine aminotransferase (ALT), an enzyme involved in Ala biosynthesis, and examined its expression profile. It was found that the full-length kuruma shrimp ALT1 cDNA consisted of 3,301 bp, encoding 514 amino acids, and that all amino acid residues important for ALT activity were conserved. Phylogenetic analysis also indicated that the ALT gene cloned in this study was classified as ALT1. Moreover, we examined the expression levels of the ALT1 gene in the abdominal muscle and the hepatopancreas of kuruma shrimp acclimated at 17‰, 34‰, and 40‰ salinities, resulting that the mRNA levels of the ALT1 genes in both tissues of the shrimp acclimated at 40‰ were significantly higher than those at 17‰ for 12 h (<i>p</i> < 0.05). The mRNA levels of the ALT1 gene in the abdominal muscle of the shrimp acclimated for more than 24 h tended to increase following the increase of environmental salinity. These results indicate that ALT1 is responsible for the increase of free Ala concentration in the abdominal muscle of kuruma shrimp to regulate osmotic pressure at high salinity.</p>\",\"PeriodicalId\":15711,\"journal\":{\"name\":\"Journal of experimental zoology. Part A, Ecological and integrative physiology\",\"volume\":\"341 5\",\"pages\":\"615-626\"},\"PeriodicalIF\":1.9000,\"publicationDate\":\"2024-03-26\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Journal of experimental zoology. Part A, Ecological and integrative physiology\",\"FirstCategoryId\":\"99\",\"ListUrlMain\":\"https://onlinelibrary.wiley.com/doi/10.1002/jez.2811\",\"RegionNum\":3,\"RegionCategory\":\"生物学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q1\",\"JCRName\":\"ZOOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of experimental zoology. Part A, Ecological and integrative physiology","FirstCategoryId":"99","ListUrlMain":"https://onlinelibrary.wiley.com/doi/10.1002/jez.2811","RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"ZOOLOGY","Score":null,"Total":0}
引用次数: 0
摘要
包括对虾在内的一些甲壳类动物会改变特定游离氨基酸的含量,以调节体内的渗透压。日本库氏对虾(Penaeus japonicus)也会随着环境盐度的升高而增加腹肌中丙氨酸(Ala)的浓度。在本研究中,为了阐明盐度对库鲁玛对虾体内丙氨酸积累变化的机制,我们克隆了参与丙氨酸生物合成的丙氨酸氨基转移酶(ALT)的编码基因,并研究了其表达谱。结果发现,全长的库氏对虾 ALT1 cDNA 由 3,301 bp 组成,编码 514 个氨基酸,而且 ALT 活性的所有重要氨基酸残基都是保守的。系统进化分析也表明,本研究克隆的 ALT 基因属于 ALT1。此外,我们还检测了在 1.7‰、3.4‰和 4.0‰盐度下驯化的库氏对虾腹肌和肝胰腺中 ALT1 基因的表达水平,结果发现在 4.0‰盐度下驯化 12 h 的库氏对虾腹肌和肝胰腺中 ALT1 基因的 mRNA 水平均显著高于在 1.7‰盐度下驯化 12 h 的库氏对虾腹肌和肝胰腺中 ALT1 基因的 mRNA 水平(p<0.05)。
Cloning and expression profile of the alanine aminotransferase gene from kuruma shrimp Penaeus japonicus exposed to different salinities
Several crustaceans including shrimps change the amount of specific free amino acids to regulate the osmotic pressure in their bodies. Kuruma shrimp Penaeus japonicus also increases the concentration of alanine (Ala) in the abdominal muscle following the increase of environmental salinity. In the present study, to elucidate the mechanisms of changes in Ala accumulation of kuruma shrimp depending on salinity, we cloned the gene encoding alanine aminotransferase (ALT), an enzyme involved in Ala biosynthesis, and examined its expression profile. It was found that the full-length kuruma shrimp ALT1 cDNA consisted of 3,301 bp, encoding 514 amino acids, and that all amino acid residues important for ALT activity were conserved. Phylogenetic analysis also indicated that the ALT gene cloned in this study was classified as ALT1. Moreover, we examined the expression levels of the ALT1 gene in the abdominal muscle and the hepatopancreas of kuruma shrimp acclimated at 17‰, 34‰, and 40‰ salinities, resulting that the mRNA levels of the ALT1 genes in both tissues of the shrimp acclimated at 40‰ were significantly higher than those at 17‰ for 12 h (p < 0.05). The mRNA levels of the ALT1 gene in the abdominal muscle of the shrimp acclimated for more than 24 h tended to increase following the increase of environmental salinity. These results indicate that ALT1 is responsible for the increase of free Ala concentration in the abdominal muscle of kuruma shrimp to regulate osmotic pressure at high salinity.
期刊介绍:
The Journal of Experimental Zoology – A publishes articles at the interface between Development, Physiology, Ecology and Evolution. Contributions that help to reveal how molecular, functional and ecological variation relate to one another are particularly welcome. The Journal publishes original research in the form of rapid communications or regular research articles, as well as perspectives and reviews on topics pertaining to the scope of the Journal. Acceptable articles are limited to studies on animals.