College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, China.
Joint International Research Laboratory of Agriculture and Agri-Product Safety of Ministry of Education of China, Yangzhou University, Yangzhou 225009, China.
Genes (Basel). 2022 Jan 22;13(2):201. doi: 10.3390/genes13020201.
Bone morphogenetic proteins (BMPs) are the structurally similar and highly conserved type of functional proteins that play an important role in hair follicle growth and development. was a differentially expressed gene in different patterns of Hu sheep lambskin identified using Agilent microarray. Since hair follicle is the basis of pattern formation of lambskin, and its growth and development is governed by dermal papilla cells (DPCs), to clarify the role of and hair follicle, our study was designed to investigate the regulation between and DPCs. Firstly, the CDS region of was cloned by 3'Race and PCR in Hu sheep and performed serious of bioinformatic analysis. Then, the effects of on DPCs were analyzed after overexpression and interference of in dermal papilla cells by CCK8, EdU, and PI assay. Additionally, qPCR was also conducted to clarify the relationship between and the TGF-β/Smad signaling pathway. A total of 1296 bp of the CDS region sequence was sucessfully cloned in Hu sheep, encoding a signal peptide of 431 amino acids, molecular weight was 49,316.9 Da and the isoelectric point (Pi) was 7.75. Nucleotide sequencing analysis of revealed that Hu sheep had high homology with and . Structure domain prediction showed that TGF-β superfamily domain exist between 330th-431th amino acid, protein is a secreted protein. In up-regulated DPCs, DPCs proliferation rate and cell cycle were significantly higher than that of NC group ( < 0.05). Meanwhile, the expression level of , , , and in TGF-β/Smad signaling pathway were significantly lower than that in NC group ( < 0.05). In down-regulated DPCs, it presented the opposite result. In conclusion, our study showed that had a positive effect on DPCs by accelerating the proliferation and cell cycle of DPCs, and hypothesized that regulate hair follicles growth and development via TGF-β/Smad signaling pathway. These findings may provide a synergistic target for the subsequent research of hair follicle growth and development.
骨形态发生蛋白（BMPs）是结构相似且高度保守的功能蛋白，在毛囊生长和发育中发挥重要作用。使用 Agilent 微阵列鉴定出在不同胡羊羔羊皮模式中差异表达的基因。由于毛囊是羔羊皮图案形成的基础，其生长和发育受真皮乳头细胞（DPCs）的控制，为了阐明的作用和毛囊，我们的研究旨在研究和 DPCs 之间的调节关系。首先，通过 3'Race 和 PCR 在胡羊中克隆的 CDS 区域，并进行了一系列生物信息学分析。然后，通过 CCK8、EdU 和 PI 测定分析过表达和干扰 DPCs 后对 DPCs 的影响。此外，还进行了 qPCR 以阐明与 TGF-β/Smad 信号通路之间的关系。成功地在胡羊中克隆了全长 1296bp 的 CDS 序列，编码 431 个氨基酸的信号肽，分子量为 49316.9Da，等电点（Pi）为 7.75。对的核苷酸序列分析表明，胡羊与和具有高度同源性。结构域预测表明，TGF-β 超家族结构域存在于 330 位-431 位氨基酸之间，蛋白是一种分泌蛋白。在上调的 DPCs 中，DPCs 的增殖率和细胞周期明显高于 NC 组（<0.05）。同时，TGF-β/Smad 信号通路中、、、的表达水平明显低于 NC 组（<0.05）。在下调的 DPCs 中，结果相反。总之，我们的研究表明通过加速 DPCs 的增殖和细胞周期，对 DPCs 有积极影响，并假设通过 TGF-β/Smad 信号通路调节毛囊的生长和发育。这些发现可能为毛囊生长和发育的后续研究提供协同靶标。
