Cloning of tight junctions protein genes Claudin-3 and Claudin-12 and their intestinal expression under the intervention of arginine in hybrid grouper Epinephelus fuscoguttatus♀×E.lanceolatus♂
LI Guang,CUI Xiao,YAO Chunfeng,CHEN Donghong,HE Kai,CHEN Guanlin,HAN Yuxuan,YANG Qihui，CHI Shuyan
1.College of Fisheries,Guangdong Ocean University,Zhanjiang 524088,China;2.Guangdong Yuehai Feeds Group Company Limited,Zhanjiang 524000,China;3.Southern Marine Science and Engineering Guangdong Laboratory(Zhanjiang),Zhanjiang 524025,China
Abstract: To evaluate the role of Claudin-3 and Claudin-12 genes in growth and intestinal mucosal barrier of hybrid grouper Epinephelusfuscoguttatus♀×E.lanceolatus♂,the full length of Claudin-3 and Claudin-12 genes of hybrid grouper with body weight of (11.0±0.5)g were cloned by homologous cloning and the expression of Claudin-3 and Claudin-12 genes was detected in brain,gill,skin,muscle,liver,stomach,foregut,midgut,hindgut,head kidney,body kidney,spleen and heart by race qRT-PCR.The expression of these two genes in intestine and growth performance were evaluated in the hybrid grouper with initial body mass of (20.79±0.09)g fed diets containing 1.96%,3.06% and 3.74% of arginine for 8 weeks at water temperature of (29±1)℃.The results showed that the Claudin-3 had full-length cDNA sequence of 1 544 bp,including a 153 bp 5′untranslated region (UTR)and a 743 bp 3′ UTR,with the open reading frame (ORF)of 648 bp encoding a predicted protein of 215 amino acids with a relative molecular mass of 23 100.The Claudin-12 had full-length cDNA sequence of 1 236 bp,including a 118 bp 5′UTR and a 92 bp 3′UTR,with the ORF of 1 026 bp encoding a predicted protein of 341 amino acids with a relative molecular mass of 37 400.Phylogenetic tree and amino acid sequence alignment revealed that Claudin-3 amino acid sequence was the closest to that of grouper Epinepheluscoioides with 98.14% homology,and Claudin-12 amino acid sequence was the closest to that of live sharksucker Echeneisnaucratus with 91.75% homology.Claudin-12 gene and Claudin-3 gene were expressed in all 13 tissues tested,significantly higher Claudin-3 mRNA expression in gill and body kidney than those in other tissues (P<0.05),followed by midgut and liver.There was the maximal expression of Claudin-12 gene in brain,significantly higher than those in other groups (P<0.05),followed by midgut,hindgut,heart,body kidney,head kidney,and spleen.Significantly higher growth was observed in the hybrid grouper fed the diet containing 3.06% arginine,than that in the other two groups (P<0.05).There were significantly higher weight grain rate and gene expression levels of Claudin-3 and Claudin-12 in the intestinal tract in the hybrid grouper fed the diet containing 3.06% arginine than those in the fish fed the diet containing 1.96% and 3.74% arginine (P<0.05),indicating Claudin-3 and Claudin-12 genes up-regulated by arginine maintaining the integrity of intestinal mucosal barrier and promoting the growth of the hybrid grouper.
李广, 崔晓, 姚春凤, 陈东鸿, 何凯, 陈冠霖, 韩煜轩, 杨奇慧, 迟淑艳. 珍珠龙胆石斑鱼紧密连接蛋白基因Claudin-3与Claudin-12的克隆及精氨酸干预下的肠道表达[J]. 大连海洋大学学报, 2022, 37(1): 61-70.
LI Guang, CUI Xiao, YAO Chunfeng, CHEN Donghong, HE Kai, CHEN Guanlin, HAN Yuxuan, YANG Qihui, CHI Shuyan. Cloning of tight junctions protein genes Claudin-3 and Claudin-12 and their intestinal expression under the intervention of arginine in hybrid grouper Epinephelus fuscoguttatus♀×E.lanceolatus♂. Journal of Dalian Ocean University, 2022, 37(1): 61-70.
2022, Vol. 37 
