Hyaluronic acid, CD44 and RHAMM regulate myoblast behavior during embryogenesis.
Hyaluronic acid, CD44 and RHAMM regulate myoblast behavior during embryogenesis.
复制标题
DOI：
10.1016/j.matbio.2018.08.008
复制
发表时间：
2019-05
期刊：
影响因子：
--
通讯作者：
Calve S
中科院分区：
文献类型：
--
作者：
Leng Y;Abdullah A;Wendt MK;Calve S
透明质酸（HA）是细胞外基质（ECM）的一种成分，研究表明它在肌肉修复与再生过程中对调节肌肉细胞行为起着重要作用。例如，肌肉损伤后的细胞外基质重塑涉及透明质酸表达上调，且这一过程与骨骼肌前体细胞的募集相关。然而，关于透明质酸在骨骼肌发育过程中的作用，人们知之甚少。为深入了解透明质酸介导胚胎期肌发生的方式，我们首先测定了胚胎第10.5天至12.5天小鼠前肢中CD44、透明质酸介导的运动受体（RHAMM）及其他与透明质酸相关蛋白的空间分布和基因表达情况。在此期间，透明质酸和CD44的表达维持在较高水平，而RHAMM无论是在蛋白质水平（通过免疫组织化学检测）还是RNA水平（通过定量聚合酶链反应检测）均有所下降。接下来，我们发现4 - 甲基伞形酮介导的透明质酸合成抑制会阻碍胚胎第11.5天/12.5天前肢来源细胞的迁移和增殖。随后，我们使用针对CD44和RHAMM这两种受体的抗体，研究它们对成肌细胞和结缔组织细胞行为的影响。抗RHAMM抗体（而非抗CD44抗体）显著减少了成肌祖细胞在24小时内迁移的总距离，而这两种抗体均抑制了结缔组织细胞的迁移。相比之下，抗CD44抗体抑制了结缔组织细胞和成肌祖细胞的增殖，而抗RHAMM抗体则无此作用。然而，当通过短发夹RNA（shRNA）使成肌细胞和结缔组织细胞中的CD44和RHAMM缺失时，这两种细胞的运动性和增殖均受到显著抑制，这表明阻断细胞表面定位的CD44和RHAMM，其效果不如通过shRNA全面敲除这些受体明显。这些结果首次揭示了RHAMM在骨骼肌环境中的分布和活性。此外，我们的数据表明，透明质酸通过与CD44和RHAMM相互作用，促进成肌祖细胞的迁移和增殖。明确透明质酸及其受体在指导肌发生中的作用，将有助于设计旨在促进受损或患病肌肉恢复的再生疗法。
Hyaluronic acid (HA) is an extracellular matrix (ECM) component that has been shown to play a significant role in regulating muscle cell behavior during repair and regeneration. For instance, ECM remodeling after muscle injury involves an upregulation in HA expression that is coupled with skeletal muscle precursor cell recruitment. However, little is known about the role of HA during skeletal muscle development. To gain insight into the way in which HA mediates embryonic myogenesis, we first determined the spatial distribution and gene expression of CD44, RHAMM and other HA related proteins in embryonic day (E)10.5 to E12.5 murine forelimbs. While HA and CD44 expression remained high, RHAMM decreased at both the protein (via immunohistochemistry) and RNA (via qPCR) levels. Next, we determined that 4-methylumbelliferone-mediated knockdown of HA synthesis inhibited the migration and proliferation of E11.5/E12.5 forelimb-derived cells. Then, the influence of CD44 and RHAMM on myoblast and connective tissue cell behavior was investigated using antibodies against these receptors. Anti-RHAMM, but not anti-CD44, significantly decreased the total distance myogenic progenitors migrated over 24 hrs, whereas both inhibited connective tissue cell migration. In contrast, anti-CD44 inhibited the proliferation of connective tissue cells and muscle progenitors, but anti-RHAMM had no effect. However, when myoblasts and connective tissue cells were depleted of CD44 and RHAMM by shRNA, motility and proliferation were significantly inhibited in both cells indicating that blocking cell surface-localized CD44 and RHAMM does not have as pronounced effect as global shRNA-mediated depletion of these receptors. These results show, for the first time, the distribution and activity of RHAMM in the context of skeletal muscle. Furthermore, our data indicate that HA, through interactions with CD44 and RHAMM, promotes myogenic progenitor migration and proliferation. Confirmation of the role of HA and its receptors in directing myogenesis will be useful for the design of regenerative therapies that aim to promote the restoration of damaged or diseased muscle.
登录
查看更多内容
影响因子：
9.7
作者：
Dicker, Kevin T.;Gurski, Lisa A.;Pradhan-Bhatt, Swati;Witt, Robert L.;Farach-Carson, Mary C.;Jia, Xinqiao
通讯作者：
Jia, Xinqiao
影响因子：
3.7
作者：
Brinck, J;Heldin, P
通讯作者：
Heldin, P
影响因子：
4
作者：
Acharya, Pinak S.;Majumdar, Sonali;Pure, Ellen
通讯作者：
Pure, Ellen
影响因子：
4.8
作者：
Bourguignon, Lilly Y. W.;Gilad, Eli;Peyrollier, Karine
通讯作者：
Peyrollier, Karine
影响因子：
5.5
作者：
Calve, Sarah;Isaac, Jahdonna;Mendias, Christopher L.
通讯作者：
Mendias, Christopher L.