Zhang Yu, Zhao Xuefei, Li Shuqi, Bai Suying, Zhang Wei
College of Wildlife and Protected Area, Northeast Forestry University, Harbin 150040, China.
National Forestry and Grassland Administration Research Center of Engineering Technology for Wildlife Conservation and Utilization, Harbin 150040, China.
Biomolecules. 2025 Jun 13;15(6):863. doi: 10.3390/biom15060863.
The development and replacement of hair play a significant role in the life history of animals. In recent years, retinoic-acid-related orphan receptor alpha () has been found to participate in the regulation of hair follicle development, yet the underlying mechanisms remain incompletely understood. This study aims to analyze the regulatory role of on the cytoskeleton of hair follicle stem cells (HFSCs). We treated HFSCs with a RORA agonist and subsequently analyzed differential gene expression using qPCR, Western blotting, and immunofluorescence, finding that agonist-induced activation of RORA suppressed the expression levels of cytoskeleton-related genes. Additionally, F-actin staining with phalloidin, followed by migration assays and wound healing tests for cell migration detection, revealed that this process affected the cytoskeletal state of HFSCs and inhibited their migration and adhesion capabilities. We further conducted interaction analyses using CUT&RUN combined with ddPCR and EMSA, demonstrating that RORA can bind to the promoter regions of the gene and regulate their transcription. This study contributes to a comprehensive understanding of the regulatory processes involved in hair follicle development and may provide broader insights into the treatment of diseases such as alopecia.
毛发的生长和更替在动物的生命历程中起着重要作用。近年来,人们发现维甲酸相关孤儿受体α(RORA)参与毛囊发育的调控,但其潜在机制仍未完全明确。本研究旨在分析RORA对毛囊干细胞(HFSCs)细胞骨架的调控作用。我们用RORA激动剂处理HFSCs,随后通过qPCR、蛋白质免疫印迹和免疫荧光分析差异基因表达,发现激动剂诱导的RORA激活抑制了细胞骨架相关基因的表达水平。此外,用鬼笔环肽进行F-肌动蛋白染色,随后进行迁移实验和伤口愈合测试以检测细胞迁移,结果表明这一过程影响了HFSCs的细胞骨架状态并抑制了它们的迁移和黏附能力。我们进一步使用CUT&RUN结合ddPCR和EMSA进行相互作用分析,证明RORA可以与该基因的启动子区域结合并调节其转录。本研究有助于全面了解毛囊发育所涉及的调控过程,并可能为脱发等疾病的治疗提供更广泛的见解。