鳜twist2序列特征、时空表达模式及靶向miRNAs预测分析OA北大核心CSTPCD

Twist2(Twist family transcription Factor 2)is a member of the basic Helix-Loop-Helix(bHLH)family that plays an important role in the regulation of animal development.However,the expression patterns and regulatory roles of twist2 genes in fishes are poorly understood.This study aimed to understand the expression pattern of the twist2 gene and its functions in the regulation of muscle development in Siniperca chuatsi.The twist2 gene sequence was obtained from the transcriptome database of S.chuatsi and Twist2 protein characterization and homology were analyzed using bioinformatic methods.The protein encoded by the twist2 gene of S.chuatsi consists of 162 amino acids with a conserved functional bHLH structural domain and the results of multiple sequence comparison based on the amino acid sequences showed that the Twist2 protein has a high similarity in vertebrates.The spatiotemporal expression pattern of the twist2 gene of S.chuatsi was analyzed by RT-qPCR and the expression pattern of the twist2 gene was found to be differentially expressed at different developmental stages.The expression level was low before the mid-gastrula stage,increased significantly from the neurula stage,reached its highest level in the tail-bud stage,and was widely expressed in S.chuatsi tissues;its expression was highest in the spleen,followed by the brain and red muscle.Whole embryo in situ hybridization was used to detect the localization of twist2 at different developmental stages in early embryos and it was found to be specifically expressed mainly in the neuroectoderm and somites.Targetscan and RNAhybrid tools were used to predict miRNAs that might target the mandarin twist2 gene.miR-30a,miR-30b,miR-30e-5p,and miR-204 were found to potentially act on the twist2 3'UTR,suggesting that twist2 is a potential target gene for miR-30a,miR-30b,miR-30e-5p,and miR-204.The short-term starvation stress experiment in S.chuatsi showed that the expression of twist2 after three days of starvation was significantly negatively correlated with the expression of the above miRNAs,suggesting that there is a potential regulatory relationship between twist2 and the above four miRNAs.Therefore,the results of this study will contribute to understanding the sequence characteristics and spatiotemporal expression patterns of the twist2 gene and the functions of potential upstream miRNAs in regulating muscle growth and development at the molecular level,which will provide a theoretical basis for the developmental biology of fish,as well as for healthy aquaculture.