Proteomics Study Of The Antler Stem Cell Tissue That Was Activated By Circulating Androgen Hormones For Antler Development

Deer antlers are not only widely known for their medicinal value,but they can also be developed into multiple novel biomedical models.Pedicle periosteum(PP)is the basis of annual regeneration of deer antlers.Its strong proliferation and differentiation potential are derived from antlerogenic periosteum(AP)that overlies each frontal crest in prepubertal deer.Both antler generation(pedicle and the first antler formation)and regeneration are regulated by androgen hormones.However,the molecular mechanisms of AP and PP development that activated by androgens are still unclear.In this study,the AP and PP tissues were taken as the research objects to get the differential expressed proteins(DEPs)that induced by androgen in each type of tissue using 2D-DIGE and Label free.According to the results of proteomics,key regulatory factors and related signaling pathways regulating the development of AP and PP were screened,and key regulatory proteins during the development of AP tissues activated by androgen were found.In addition,we also explored whether androgens could promote proliferation of the AP cells when the AP tissues are cultured in vitro.The results were shown that 83 DEPs(15 up-regulated and 68 up-regulated),identified using 2D-DIGE in the AP development induced by androgens,were mainly involving cell cycle,programmed apoptosis,gene expression and signal transduction.GO functional analysis revealed that DEPs were clustered in biological processes such as protein nucleus localization,neuron regeneration and cytoskeleton reorganization,and important regulatory factors such as CALR,p53 and SRCI1 were screened for AP tissue development.The CALR,p53,LMNA and ACTB were involved in both antler generation and regeneration.In addition,CALR,p53 and ACTB were expressed in opposite levels in both processes.During the development and regeneration of antler,CALR and p53 could interact directly with AR,and their expression levels were closely associated with androgen levels.The results of EdU labeling and Ki67 immunohistochemistry showed that androgen could not promote proliferation of the AP cells when the AP tissues were cultured in vitro.Conclusion: 1)High level of androgen is the key factor to activate AP development,and low level of androgen is the key factor to activate PP development;2)The development of AP is mainly regulated by cell cycle,programmed cell apoptosis,gene expression and signal transduction,and CALR,p53 and SRCI1 are the key regulatory genes involved in AP development;3)The development of PP is mainly regulated by endoplasmic reticulum protein.Heat shock protein,CALR,FK506 and SRTK are the main factors regulating the development of PP.4)Androgen has no effect on the proliferation of AP cells when the AP tissues were cultured in vitro.