Department of Cell and Developmental Biology, Sackler Faculty of Medicine, Tel-Aviv University, Tel-Aviv, 69978, Israel.
Stem Cell Rev Rep. 2010 Mar;6(1):42-9. doi: 10.1007/s12015-009-9106-9.
In this study we explored the expression pattern of SVEP1, a novel cell adhesion molecule (CAM), in bona fide satellite cells and their immediate progeny. We show that SVEP1 is expressed in activated satellite cells prior to their determination to the myogenic lineage. SVEP1 was also expressed during early phases of myogenic differentiation through the initial stage of myoblast fusion and myotube formation. The expression of SVEP1 was shown by immunostaining two cell culture systems: freshly isolated myofibers and primary myoblasts. Pax7 was used to pinpoint satellite cells situated in their niche on myofibers, and activated satellite cells were determined based on BrdU incorporation (Pax7(+)/BrdU(+)cells). MyoD marked satellite cells fated to undergo myogenesis as well as proliferating and differentiating myoblasts. Differentiating myoblasts and myotubes were identified based on their sarcomeric myosin expression. We showed that SVEP1 was specifically expressed in pre-determined activated satellite cells (Pax7(+)/ BrdU(+) /MyoD(-)) accounting for about 24% of total satellite cells. On the other hand, SVEP1 expression was absent in quiescent satellite cells (Pax7(+)/BrdU(-)/MyoD(-)). Moreover, based on MyoD/sarcomeric myosin co-expression SVEP1 was shown to be expressed throughout the early phases of myogenesis up until myoblast fusion and myotube formation. A decline in SVEP1 expression occurred upon myotube maturation. We suggest SVEP1 as a potential biomarker for pre-fated satellite cells. The impact of this finding is that it may allow scrutinizing signals that affect differentiation commitment. Thus, holds a therapeutic potential for maladies that involve deregulated stem cell fate-decision.
在这项研究中,我们探索了 SVEP1 的表达模式,SVEP1 是一种新的细胞粘附分子 (CAM),存在于真正的卫星细胞及其直接祖细胞中。我们表明,SVEP1 在卫星细胞被决定向肌生成谱系之前就已经在激活的卫星细胞中表达。SVEP1 在肌生成分化的早期阶段,即在成肌细胞融合和肌管形成的初始阶段也有表达。免疫染色两种细胞培养系统显示了 SVEP1 的表达:新鲜分离的肌纤维和原代成肌细胞。使用 Pax7 标记位于肌纤维龛位的卫星细胞,基于 BrdU 掺入来确定激活的卫星细胞(Pax7(+)/BrdU(+) 细胞)。MyoD 标记卫星细胞注定要进行肌生成以及增殖和分化的成肌细胞。基于肌球蛋白的表达来鉴定分化的成肌细胞和肌管。我们表明,SVEP1 特异性表达在预先确定的激活的卫星细胞(Pax7(+)/BrdU(+) /MyoD(-))中,占总卫星细胞的约 24%。另一方面,SVEP1 表达在静止的卫星细胞(Pax7(+)/BrdU(-)/MyoD(-))中不存在。此外,基于 MyoD/肌球蛋白的共表达,SVEP1 被证明在肌生成的早期阶段表达,直到成肌细胞融合和肌管形成。SVEP1 的表达在肌管成熟时下降。我们认为 SVEP1 是预先确定的卫星细胞的潜在生物标志物。这一发现的影响是,它可能允许仔细研究影响分化承诺的信号。因此,它在涉及失调的干细胞命运决策的疾病中具有治疗潜力。