MOLECULAR AND CELLULAR NUTRITION

Clone of Zonula Occluden-1, Occludin and Claudin-15a Genes from Hybrid Grouper (Epinephelus fuscoguttatus♀×Epinephelus lanceolatus) and Its Expression in Intestinal Tissue under Intervention of Arginine

  • CHEN Donghong ,
  • CUI Xiao ,
  • LI Guang ,
  • HE Kai ,
  • HAN Yuxuan ,
  • CHEN Guanlin ,
  • YANG Qihui ,
  • CHI Shuyan
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  • 1. College of Fisheries, Guangdong Ocean University, Zhanjiang 524088, China;
    2. Guangdong Laboratory of Southern Marine Science and Engineering (Zhanjiang), Zhanjiang 524025, China

Received date: 2021-01-18

  Online published: 2021-08-11

Supported by

 

Abstract

This experiment was conducted to study the clone of Zonula Occluden-1 (ZO-1), Occludin and Claudin-15a genes from hybrid grouper (Epinephelus fuscoguttatus♀×Epinephelus lanceolatus) and its expression in intestinal tissue under intervention of arginine. The full-length cDNAs of ZO-1, Occludin and Claudin-15a of hybrid grouper were cloned by rapid amplification of cDNA ends (RACE)-PCR technology, and the mRNA relative expression levels of 13 kinds of tissues (brain, gill, skin, muscle, liver, stomach, fore-intestine, mid-intestine, hind-intestine, head kidney, body kidney, spleen and heart) of hybrid grouper were analyzed. A total of 375 hybrid grouper with similar body weight of (20.79±0.09) g were randomly divided into 3 groups with 5 replicates per group and 25 fish per replicate. The dietary arginine levels of each groups were 1.96%, 3.06% and 3.74%, respectively. Feeding for 8 weeks, and the mRNA relative expression levels of ZO-1, Occludin and Claudin-15a in intestine were measured. The results showed as follows:1) the full-length cDNA of ZO-1 consisted of 6 355 bp, contained an open reading frame (ORF) of 5 106 bp, encoded 1 701 amino acids, contained a 105 bp 5'-untranslated region (UTR) and a 1 144 bp 3'-UTR, and had no transmembrane region. The full-length cDNA of Occludin consisted of 2 222 bp, contained an ORF 1 515 bp, encoded 504 amino acids, contained a 69 bp 5'-UTR and a 638 bp 3'-UTR, and had 4 transmembrane regions. The full-length cDNA of Claudin-15a consisted of 1 584 bp, contained an ORF 669 bp, encoded 222 amino acids, contained a 387 bp 5'-UTR and a 528 bp 3'-UTR, and had 5 transmembrane regions. 2) The phylogenetic tree analysis and amino acid sequence alignment showed that ZO-1 of hybrid grouper had the highest homology with Seriola dumerili (94%), the Occludin had the highest homology with Larimichthys crocea (84%) and the Claudin-15a had the highest homology with Sphaeramia orbicularis (92%). 3) The mRNA expression relative level of ZO-1 in hind-intestine was significantly higher than that of other tissues (P<0.05), while the mRNA expression relative level of ZO-1 in brain, gills, heart, head kidney, stomach, liver, muscle and body kidney was no significant difference (P>0.05); the mRNA expression relative level of Claudin-15a in fore-intestine and mid-intestine was significantly higher than that of other tissues (P<0.05); the mRNA expression relative level of Occludin in skin, fore-intestine, mid-intestine, hind-intestine and gill was significantly higher than that of other tissues (P<0.05). 4) The weight gain rate and mRNA expression relative levels of ZO-1, Claudin-15a and Occludin in intestine of 3.06% arginine level group were significantly higher than those of 1.69% and 3.74% arginine level groups (P<0.05). In conclusion, the full-length cDNAs of ZO-1, Occludin and Claudin-15a of hybrid grouper are cloned by RACE-PCR technology in this experiment. Dietary appropriate arginine level can significant improve the mRNA expression relative levels of ZO-1, Claudin-15a and Occludin in intestine of hybrid grouper, maintain the intestinal mucosal barrier integrity, and promote the healthy growth of fish.

Cite this article

CHEN Donghong , CUI Xiao , LI Guang , HE Kai , HAN Yuxuan , CHEN Guanlin , YANG Qihui , CHI Shuyan . Clone of Zonula Occluden-1, Occludin and Claudin-15a Genes from Hybrid Grouper (Epinephelus fuscoguttatus♀×Epinephelus lanceolatus) and Its Expression in Intestinal Tissue under Intervention of Arginine[J]. Chinese Journal of Animal Nutrition, 2021 , 33(8) : 4645 -4661 . DOI: 10.3969/j.issn.1006-267x.2021.08.044

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