Neural cell adhesion molecule (NCAM) marks adult myogenic cells committed to differentiation
Neural cell adhesion molecule (NCAM) marks adult myogenic cells committed to differentiation
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DOI：
10.1016/j.yexcr.2008.01.021
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发表时间：
2008-04-15
影响因子：
3.7
通讯作者：
Cornelison, D. D. W.
中科院分区：
文献类型：
--
作者：
Capkovic, Katie L.;Stevenson, Severin;Cornelison, D. D. W.
虽然最近在大规模基因表达分析方面的进展大大增加了我们对多种细胞类型中存在的mRNA库的了解，但越来越清楚的是，对编码蛋白的表达、定位和关联的检查对于确定其功能意义至关重要。特别是已经提出许多信号受体、转换器和效应器以与质膜的物理上不同的区域相关的更高级复合物的形式起作用。成体肌肉干细胞（卫星细胞）必须在损伤后对广泛的细胞外刺激做出适当的反应：因此，这种信号支架的作用是一个潜在的重要研究领域。为了解决这个问题，我们首先从原代卫星细胞中分离出耐洗涤剂的膜组分，然后使用液相色谱-串联质谱法分析其组分蛋白。粘附介导的信号通路的跨膜和跨膜组分构成了最大的一组已鉴定的蛋白质;特别是神经细胞粘附分子（NCAM），一种以前与肌肉再生相关的多功能细胞表面蛋白，具有重要意义。免疫组织化学分析显示，NCAM不仅定位于质膜的离散区域，它也是一个非常早期的终末分化的标志物。使用流式细胞术，我们已经分选出物理上同质的肌源性培养物，仅根据NCAM表达分为增殖和分化部分。(C)2008年爱思唯尔公司All rights reserved.
Although recent advances in broad-scale gene expression analysis have dramatically increased our knowledge of the repertoire of mRNAs present in multiple cell types, it has become increasingly clear that examination of the expression, localization, and associations of the encoded proteins will be critical for determining their functional significance. in particular, many signaling receptors, transducers, and effectors have been proposed to act in higher-order complexes associated with physically distinct areas of the plasma membrane. Adult muscle stem cells (satellite cells) must, upon injury, respond appropriately to a wide range of extracellular stimuli: the role of such signaling scaffolds is therefore a potentially important area of inquiry. To address this question, we first isolated detergent-resistant membrane fractions from primary satellite cells, then analyzed their component proteins using liquid chromatography-tandem mass spectrometry. Transmembrane and juxtamembrane components of adhesion-mediated signaling pathways made up the largest group of identified proteins; in particular, neural cell adhesion molecule (NCAM), a multifunctional cell-surface protein that has previously been associated with muscle regeneration, was significant. Immunohistochemical analysis revealed that not only is NCAM localized to discrete areas of the plasma membrane, it is also a very early marker of commitment to terminal differentiation. Using flow cytometry, we have sorted physically homogeneous myogenic cultures into proliferating and differentiating fractions based solely upon NCAM expression. (C) 2008 Elsevier Inc. All rights reserved.