The Role And Mechanism Of Arctic Fox Wnt3a Gene In The Process Of Melanin Formation

Arctic fox fur is bright in color and thick in fur,has excellent thermal insulation properties,and plays an important role in fur trading.The hair color has a huge impact on fur quality,so research on the formation mechanism of hair color has become a hot spot in recent years.However,there are few reports on the formation mechanism of arctic fox fur.I have analyzed the arctic fox embryonic transcriptome analysis and found that the Wnt3 a gene has an important role in the formation of melanin,but research involving the arctic fox has never been found.Therefore,the role and mechanism of the arctic fox Wnt3 a gene in the formation of melanin are studied in this article,with a view to further theoretically supplement and pave the way for further exploration of the mechanism of arctic fox hair color formation.In this study,gene overexpression technology was used to construct an overexpression vector of Arctic fox Wnt3 a gene.After transfection into A375 cells,the expression levels of Wnt3 a,MITF,TYR,TYRP1,and PMEL genes were detected by real-time quantitative PCR.At the same time,Western blot was used to detect the expression of TYRP1 and PMEL proteins.In addition,reports have found that Wnt3 a regulates melanin formation through the MITF gene.Therefore,this study also investigated the association between MITF and its downstream genes.An overexpression vector of Arctic fox MITF gene was constructed,and after transfection into A375 cells,the expression levels of its downstream genes TYR,TYRP1,and PMEL were measured.At the same time,according to the firefly luciferase reporter gene expression vector constructed by the research team and containing the arctic fox TYR and TYRP1 gene promoter fragments,the binding relationship between them and the MITF gene was studied using the double luciferase detection technology.The results showed that:1.Arctic fox Wnt3 a gene overexpression can regulate the expression of melanin by promoting the expression of MITF,TYR,TYRP1,and PMEL genes.2.Arctic fox MITF gene overexpression can promote the expression of downstream genes TYR,TYRP1 and PMEL.3.There is a MITF binding site in the promoter region of the Arctic fox TYR gene,which is located in the promoter region-90/-89 region.It can positively regulate the transcription of the Arctic fox TYR gene.4.There is a MITF binding site in the promoter region of the Arctic Fox TYRP1 gene,which is located in the promoter region-747/-746.It can positively regulate the transcription of the Arctic Fox TYRP1 gene.