The Laboratory of Cell and Developmental Biology, Northeast Agricultural University, Harbin, Heilongjiang 150030, China.
The Laboratory of Cell and Developmental Biology, Northeast Agricultural University, Harbin, Heilongjiang 150030, China.
Exp Cell Res. 2018 May 15;366(2):199-204. doi: 10.1016/j.yexcr.2018.03.027. Epub 2018 Mar 22.
C2C12 murine myoblasts are a common model for studying muscle differentiation. Platelet endothelial aggregation receptor-1 (PEAR1), an epidermal growth factor repeat-containing transmembrane receptor, is known to participate in platelet contact-induced activation. In the present study, we demonstrated that PEAR1 is involved in the differentiation of C2C12 murine myoblasts. Western blotting and immunofluorescence staining were used to determine PEAR1 expression and localization during C2C12 cell differentiation. Subsequently, PEAR1 expression was activated and inhibited using clustered regularly interspaced short palindromic repeats-dCas9 technology to explore its effects on this process. PEAR1 expression was found to increase over the course of C2C12 cell differentiation. This protein was predominately localized on the membrane of these cells, where it clustered upon induction of differentiation. Expression of the myogenic markers Desmin, MYOG, and MYH2 revealed that PEAR1 positively regulated C2C12 cell differentiation. Moreover, induction of muscle injury by administration of bupivacaine to mice indicated that PEAR1 might play a role in muscle regeneration. In summary, our study confirmed the involvement of PEAR1 in C2C12 cell differentiation, contributing to our understanding of the molecular mechanisms underlying muscle development.
C2C12 鼠肌母细胞是研究肌肉分化的常用模型。血小板内皮细胞聚集受体-1(PEAR1)是一种含有表皮生长因子重复的跨膜受体,已知参与血小板接触诱导的激活。在本研究中,我们证明了 PEAR1 参与 C2C12 鼠肌母细胞的分化。使用 Western blot 和免疫荧光染色来确定 PEAR1 在 C2C12 细胞分化过程中的表达和定位。随后,使用簇状规则间隔短回文重复序列-dCas9 技术激活和抑制 PEAR1 表达,以探讨其对该过程的影响。研究发现,PEAR1 的表达在 C2C12 细胞分化过程中增加。这种蛋白质主要定位于这些细胞的膜上,在诱导分化时会聚集。肌球蛋白标志物 Desmin、MYOG 和 MYH2 的表达表明 PEAR1 正向调节 C2C12 细胞分化。此外,给小鼠注射布比卡因诱导肌肉损伤表明 PEAR1 可能在肌肉再生中发挥作用。总之,我们的研究证实了 PEAR1 参与 C2C12 细胞分化,有助于我们理解肌肉发育的分子机制。