The Effect Of Androgen On Follicles And Keratin Production In Hetian Sheep

[Objective] To investigate the optimal androgen concentration for culturing Hetian sheep hair follicle and to detect effects of androgen concentration on hair follicle cell proliferation and apoptosis we using immunofluorescence labeling and real-time quantitative fluorescence determinations of wool keratin-associated protein gene expression levels.[Methods] Hair follicles were isolated by microdissection and wool follicles and skin pieces were cultured in various concentrations of dihydrotestosterone(DHT)in culture medium.Next,daily lengthwise growth measurements of hair follicles were obtained using a microscopic micrometer.Cultured Hetian wool follicles were stained using the SACPIC method to reveal hair follicle structure,while sheepskin slices were used to observe cell proliferation by immunostaining and cell apoptosis using the TUNEL method.At the molecular biological level,keratin-associated protein(Kap)gene expression was studied using hair follicles cultured for various numbers of days in vitro.[Results]Effects of androgen concentrations on Hetian wool follicle growth and development were experimentally studied.EdU proliferation assays revealed that androgen promoted cell proliferation within hair follicle dermal papillae.TUNEL apoptosis detection demonstrated that androgen treatment could delay cell apoptosis.qPCR results showed that Kap1.1,Kif1.2,Kap2.12,Kap4.2,Kap7 and Kap8 expression differences were most significant in mountain type of Hetian sheep;Kap2.9,Kap6.1 and Kap5.5 is significant in plain typeof Hetian sheep,while Kap3.2 has no significant difference in the mountainous peace prototype.[Conclusion]The concentration of androgen at concentration of 100 nM can promote the growth of Hetian wool sacfollicle cells and over-expressionin vitro,resulting in overexpression of some genes of the Kap and Kif gene family.