Structure, Expression Pattern And Function Analysis Of SoxD Subfamily In Yellow River Carp(Cyprinus Carpio Var)

The Sox gene family is a kind of transcription factor which associated with mammalian testes decided gene,widely exists in the whole biosphere and participate in the multiple development of biological processes.SoxD belongs to the Sox gene family,which has been confirmed closely related to the development of the nervous system,and are expressed in the neural stem cells and the development of biological neurons,but its research in freshwater fish are less.In this study,we were able to gain three SoxD sequences from transcriptome sequencing data of Cyprinus carpio constructed by our laboratory:CcSox5,CcSox6 and CcSox13?Cyprinus carpio SoxD?.The full-length sequences of these genes were verified by PCR sequencing,blast and homology alignment,then the gene sequence structure were analyzed by bioinformatics methods.The results showed that:the full-length sequences of these three genes are 2800bp,3000bp,2366bp and encode a 780,784 and 602 amino acid protein,respectively.CcSox5 and CcSox6 contains two exons and 1 intron,CcSox13 contains four exons,3 introns.Chromosome synteny analyses revealed that CcSox5 and CcSox13 were tightly linked with the etnk gene,which was conserved in all species;however,there were no conserved regions flanking CcSox6.Numerous essential transcription factor binding sites?TFs?were predicted within the 2000 bp upstream of the 5'end of these genes.These TFs include BSX,BRN4 and NGN–NEUROD,which have been shown to be involved in the early stages of neuronal determination and neurogenesis in vertebrates.This suggest that the function of these three genes may be related to the development of neurogenesis in the Yellow River.Location and tissue distribution analyses of these three genes in different tissues and different stages of embryonic development by Quantitative real-time RT-PCR?qRT-PCR?and in situ hybridization?ISH?.It revealed that CcSox5 were mainly expressed in the brain,and the expression level was low in other tissues.The expression of CcSox5 is significantly increased in neural embryo period of embryonic development;the expression of CcSox6 and CcSox13 is also particular high in the brain,in other tissues such as muscle,eye,heart,spleen,is expressed common and present a certain gender dimorphism,shows they are not only related to brain development,but also gender development.Expression of CcSox6 is low in embryo,CcSox13 expression is very high at the early stages of the embryo but in the tail bud period is decreased.Observing the development processing of Yellow River carp embryos,larvae and young fish of after exposure of?AgNP?silver nanoparticles and silver nitrate?AgNO₃?solution.The results showed that the exposure can make multiple types of deformity of embryogenesis,and with the increase of treatment concentration,embryo hatchability reduced,deformity rate increases,higher mortality;Silver also rise as the concentration increases mortality;detecting the expression level of CcSoxD in the different development stages of the embryo and larvae of different tissues after treatment.Result shows that with the increase of the concentration,there is a decrease in the expression of the trend,nano silver and silver nitrate on the growth of fish and neural development can cause certain effect.In conclusion,we hypothesized that CcSox5,CcSox6,CcSox13 is conservative in the evolution, and may play important role in the process of embryonic development,and participate in carp neural development and the development process.