Comparative Analysis Of Transcriptome Of Musk Gland And Skin And Localization Of Expression Of Sebaceous Gland Regulatory Factors

Forest musk deer is a national first-class endangered wild animal.The musk secreted from the musk gland of mature male individuals is an important Chinese medicine raw material in the traditional Chinese medicine industry,and it is also a widely used perfume material.Musk has high medical value,so it has a pivotal position in many industries.In recent years,the scale of the breeding of forest musk deer has expanded,but the related research on forest musk deer,especially on the secretion mechanism of musk,is still lagging behind.Studies have shown that the musk gland of forest musk deer contains sebaceous glands,and the sebum secreted by them is an important component of musk.Other studies have shown that SOX9,Cav1,and AR participate in the regulation of sebum secretion in the sebaceous glands of the skin.Therefore,the primary purpose of this study is to explore whether the sebaceous gland secretion regulators,SOX9,Cav1,and AR,can be expressed and located in the musk glands and whether they play roles in the musk secretion process,to provide theoretical support for improving the production and quality of musk.In order to explore whether there is the expression of sebaceous gland secretion regulators in the musk gland,this study performed transcriptome sequencing on the skin and musk gland tissue of forest musk deer and used bioinformatics analysis methods to predict the basic information of SOX9,Cav1,and AR.Through molecular biology experiments such as RT-qPCR,IHC,and Western blotting,the expression level of genes and the tissue localization of proteins were explored,and the following main results were obtained:(1)Transcriptome sequencing analysis of the skin and musk gland tissues of the forest musk deer showed that there are genes related to the regulation of sebaceous gland function in the musk gland.(2)The homology of SOX9,Cav1,and AR and the physicochemical property,functional structure,modification site,subcellular location,and secondary and tertiary structure of proteins were analyzed by bioinformatics methods.(3)Molecular biology experiments such as RT-qPCR,IHC,and Western blotting showed that SOX9,Cav1,and AR were expressed and localized in the back and abdominal skin and musk gland tissue.The RT-qPCR results were highly consistent with the transcriptome results,indicating the reliability of the transcriptome data.The results of IHC and Western blotting showed that SOX9,Cav1,and AR were expressed in the musk gland and skin.And SOX9 and AR were located in the nucleus,and Cav1 was located in the cell membrane.(4)RT-qPCR results showed that the expression levels of SOX9,Cav1,and AR genes in the abdominal skin were significantly higher than those in the back skin and musk gland.The expression levels of SOX9,Cav1,and AR genes in the back skin and musk glands were not significantly different;IHC and Western blotting results showed that the expression levels of SOX9,Cav1,and AR proteins in the back skin and abdominal skin were similar,and they were all lower than those in the musk gland.(5)The results of HE staining showed for the first time the microstructure of hair follicles and sebaceous glands in the skin and the scent glands in musk glands.In summary,this study used bioinformatics analysis and molecular experiments to find that the musk gland of forest musk deer has the same expression of sebaceous gland regulatory factors as the skin,indicating that SOX9,Cav1,and AR have the potential to regulate the secretion function of musk gland.The results of this study provided valuable scientific information for further research on the secretion mechanism of forest musk deer and new research ideas for improving the production and quality of musk.