Regulation Of MAP3K1 On Hair Follicle Stem Cells Development Of The Yangtze River Delta White Goat

The Yangtze River Delta White Goat is a local breed in Jiangsu with excellent skin,meat and hair,and it is also the only goat breed in the world that can produce superior-quality brush hair.As a raw material for making high-grade writing brushes,superior-quality brush hair has high economic value.The previous research team used high-throughput transcriptome to sequence the goat skin tissues of superior-quality brush hair and normal-quality brush hair goats in the Yangtze River Delta and found that MAP3K1 is differentially expressed in the skin tissue of superior-quality brush hair goats.The development of hair follicle stem cells is very important for hair growth,and the molecular mechanism by which MAP3K1 regulates the development of hair follicle stem cells is still unclear.In this study,relevant molecular biological experimental techniques were used to explore the regulatory effect of MAP3K1 on hair follicle stem cells and transcriptome analysis.The experimental process and specific results were as follows:1.Tissue expression profile analysis.The expression levels of MAP3K1 in different tissues of white goats in the Yangtze River Delta were investigated by RT-qPCR,and the results of transcriptome sequencing were verified at the same time.The results showed that MAP3K1 gene was widely expressed in different tissues of goats,and the expression in spleen,lung,testis and skin tissue was higher than that in heart tissue.By comparing the expression level of MAP3K1 in the skin tissue of the neck and ridge,it was found that the expression level of MAP3K1 in superior-quality brush hair type goats was significantly higher than that of normal-quality brush hair type goats(P<0.01),in line with the previous sequencing results2.Interfering with the MAP3K1 gene.The interfering adenovirus vector was constructed,and the effects of interfering MAP3K1 on the proliferation and apoptosis of hair follicle stem cells were investigated by RT-qPCR,Western blot,CCK-8,EDU,cell cycle and apoptosis tests.The results showed that the interference efficiency of MAP3K1-sh2 on adenovirus vectors reached about 70%(P<0.01);The proliferation ability of hair follicle stem cells was significantly decreased after interference with MAP3K1(P<0.01),the number of cells in S phase was significantly increased,indicating that the cells were arrested in S phase;The apoptosis rate of hair follicle stem cells was significantly increased after interference with MAP3K1(P<0.01).These results suggested that interference with MAP3K1 can inhibit cell proliferation and promote cell apoptosis in hair follicle stem cells.3.Overexpression of MAP3K1 gene.The overexpression vector was constructed,and the effects of overexpression of MAP3K1 on the proliferation and apoptosis of hair follicle stem cells were investigated by RT-qPCR,Western blot,CCK-8,EDU,cell cycle and apoptosis tests.The results showed that the expression of MAP3K1 in the overexpression group was 10 times higher than that in the control group(P<0.01);Overexpression of MAP3K1 significantly increased the proliferation of hair follicle stem cells(P<0.01),the number of S phase cells was significantly decreased(P<0,01);The apoptotic rate of hair follicle stem cells significantly decreased after MAP3K1 overexpression(P<0.01).These results suggested that overexpression of MAP3K1 promoted the proliferation and inhibited the apoptosis of hair follicle stem cells.4.Transcriptomics analysis successfully constructed 9 mRNA libraries for interference with MAP3K1,overexpression of MAP3K1 and control group.GO and KEGG analysis show that,3913 differential genes were found,of which 2213 were up-regulated and 1700 were down-regulated.These differential genes are related to TNF,MAPK,and NOD-like receptor signaling pathways,and then reveal the regulatory molecular mechanism of MAP3K1 on hair follicle stem cells.In summary,MAP3K1 can effectively regulate the proliferation and apoptosis process of hair follicle stem cells of Yangtze River Delta white goats,and its regulatory mechanism is related to TNF,MAPK,and NOD-like receptor signaling pathways.This study provides a theoretical basis for further exploring the formation mechanism of superior-quality brush hair traits of Yangtze River Delta White Goats.