Role Of MicroRNA-1 In The Regulation Of Zebrafish Craniofacial Development

MicroRNAs(miRNAs)are endogenous non-coding single strand small molecule RNAs,with a length of approximately 22 nucleotides,which play a vital role in the development of embryo by promoting the degradation of target gene mRNA or inhibiting its translation,thus realizing the regulation of post gene transcription.Malformation caused by craniofacial dysplasia is a common type of birth defects.Neural crest cells are one of the main sources of craniofacial development cells.Most of the soft and hard tissues involved in craniofacial formation are formed.The abnormalities of any part of the neural crest cells in the process of formation,proliferation,migration and differentiation will lead to craniofacial defect.The gene and signal transduction pathway of zebrafish and human in the process of craniofacial development are highly conservative.As a model animal,it has the advantages of high reproductive efficiency,short growth cycle,transparent embryo and convenient gene operation in the development of genetic biology.This makes zebrafish become an ideal experimental animal in the development mechanism and the function of miRNA.Studies have shown that miR-1 plays an important role in the development and disease of the cardiac and skeletal muscle,However,its roles in neural crest cells induction,migration and derivation are not clear.In this study,we knock down the expression of miR-1 in zebrafish with miR-1 morpholino.We find defects in the tissues derived from the neural crest cells:a severely reduced mandible and delayed appearance of pigment cells.At 24 hpf,the reduced expression of msxb,dlx2,dlx3b,snail1b,tfap2a,ngnl and crestin indicate that miR-1 deficiency affects neural crest cells migration and differentiation.In addition,the knockdown of miR-1 gene affects the proliferation,apoptosis of neural crest cells.To provide insights into the mechanism of miR-1,we found that sec63 is a direct target gene for miR-1 through differential proteomics study(iTRAQ)and bioinformatics analysis.The injection of miR-1 antisense oligonucleotides and sec63 MO into zebrafish embryos can partly rescue embryo development abnormalities caused by knocking down miR-1.In summary,this study shows that miR-1 participates in the regulation of neural crest cell development.In addition,it acts,at least partially,by regulating sec63 expression.