| Cashmere produced by cashmere goats is a high-quality textile raw material and an economic product that is in great demand in people’s lives.The hair follicle is the main unit that produces a single visible hair shaft,its quantity and characteristics of hair follicles affect the yield and quality of cashmere.Therefore,it is necessary to study the related regulation mechanism of the morphogenesis and development of cashmere goat hair follicles.The morphogenesis and development of hair follicles is a complex process which depends on the interaction and signal communication between epidermal and dermal cells.This process is regulated by a variety of factors and is accompanied by the activation and silencing of many genes,in which transcription factors play an important role in regulating gene expression.It has evidenced that the transcription factor JUNB is involved in the morphogenesis and development of hair follicles,but there are few studies on the function of this transcription factor in hair follicle cells and related regulatory mechanisms.Therefore,based on the transcriptome sequencing data from the skin tissue of Shanbei white cashmere goats at different developmental stages,the function and regulation mechanism of transcription factor JUNB on the development of hair follicles were explored.The main results were as follows:1.The transcriptome sequencing data from the skin tissues were analyzed in the embryonic stage 65 d and 120 d embryonic stage of Shanbei white cashmere goats,and the differentially expressed gene JUNB was screened out.The results of q RT-PCR showed that JUNB was highly expressed in the embryonic stage at 120 days and low in the embryonic stage at 65 days,which was consistent with the sequencing results.Immunofluorescence revealed that JUNB was mainly expressed at 120 days of embryonic stage,which further verified the sequencing results.2.JUNB overexpression adenovirus was packaged to infect primary hair papilla cells.MTT and CCK8 experiments showed that the viability of dermal papilla cells increased after the overexpression of JUNB compared with the negative control(P<0.05).Further,through the Ed U experiment,it was found that the number of positive cells in the JUNB overexpression group increased compared with the control group(P<0.01).The above results indicateed that the transcription factor JUNB could promote the proliferation of dermal papilla cells.3.The target gene of the transcription factor JUNB was predicted by JASPAR,TRRUST and other software,and TCF3 were screened out.The results of dual luciferase experiment and q RT-PCR showed that the transcription factor JUNB targeted and up-regulated TCF3.TCF3 is a member of the TCF family in the Wnt/β-catenin signaling pathway.The results of q RT-PCR and Western Blot showed that the expression of the key gene β-catenin in the Wnt/β-catenin signaling pathway was significantly increased after the overexpression of JUNB,and the expression of its important gene LEF1 and downstream targets cyclin D1 and c-myc also increased significantly.The above results indicated that the transcription factor JUNB enhanced the activity of the Wnt/β-catenin signaling pathway and promotes the expression of genes related to cell proliferation.In summary,this study revealed the function of transcription factor JUNB in the proliferation of papilla cells of Shanbei white cashmere goats,and clarified that the transcription factor JUNB promotes the expression of cell proliferation related genes by enhancing the activity of the Wnt/β-catenin signaling pathway,and thus promotes the proliferation of hair nipple cells.This study provides a certain theoretical basis for the molecular regulation on hair morphogenesis and development,and further has important theoretical significance for improving the yield and quality of cashmere goats. |
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