Dominov J A, Houlihan-Kawamoto C A, Swap C J, Miller J B
Myogenesis Research Laboratory, Massachusetts General Hospital, Charlestown, Massachusetts 02129, USA.
Dev Dyn. 2001 Jan;220(1):18-26. doi: 10.1002/1097-0177(2000)9999:9999<::AID-DVDY1088>3.0.CO;2-#.
Apoptotic myonuclei appear during myogenesis and in diseased muscles. To investigate cell death regulation in skeletal muscle, we examined how members of the Bcl-2 family of apoptosis regulators are expressed and function in the C2C12 muscle cell line and in primary muscle cells at different stages of development. Both anti-apoptotic (Bcl-W, Bcl-X(L)) and pro-apoptotic (Bad, Bak, Bax) members of the Bcl-2 family were expressed in developing skeletal muscle in vivo. Each was also expressed in embryonic (E11-12), fetal (E15-16), and neonatal muscle stem cells, myoblasts, and myotubes in vitro. In contrast, Bcl-2 expression was limited to a small group of mononucleate, desmin-positive, myogenin-negative muscle cells that were seen in fetal and neonatal, but not embryonic, muscle cell cultures. The cell surface protein Sca-1, which is associated with muscle and blood stem cells, was found on approximately 1/2 of these Bcl-2-positive cells. Loss of Bcl-2 did not affect expression of other family members, because neonatal muscles of wild-type and Bcl-2-null mice had similar amounts of Bcl-X(L), Bcl-W, Bad, Bak, and Bax mRNAs. Loss of Bcl-2 did have functional consequences; however, because neonatal muscles of Bcl-2-null mice had only approximately 2/3 as many fast muscle fibers as muscles in wild-type mice. Thus, Bcl-2 function is required for particular stages of fetal and postnatal myogenesis.
凋亡性肌细胞核在肌生成过程以及患病肌肉中出现。为了研究骨骼肌中的细胞死亡调控,我们检测了凋亡调节因子Bcl-2家族成员在C2C12肌细胞系以及处于不同发育阶段的原代肌细胞中的表达情况和功能。Bcl-2家族的抗凋亡成员(Bcl-W、Bcl-X(L))和促凋亡成员(Bad、Bak、Bax)在体内发育中的骨骼肌中均有表达。它们在体外的胚胎(E11-12)、胎儿(E15-16)和新生肌干细胞、成肌细胞以及肌管中也均有表达。相比之下,Bcl-2的表达仅限于一小群单核、结蛋白阳性、肌细胞生成素阴性的肌细胞,这些细胞可见于胎儿和新生肌细胞培养物中,但不见于胚胎肌细胞培养物中。细胞表面蛋白Sca-1与肌肉和血液干细胞相关,在大约一半的这些Bcl-2阳性细胞上被发现。Bcl-2的缺失并不影响其他家族成员的表达,因为野生型和Bcl-2基因敲除小鼠的新生肌肉中Bcl-X(L)、Bcl-W、Bad、Bak和Bax mRNA的含量相似。然而,Bcl-2的缺失确实产生了功能后果,因为Bcl-2基因敲除小鼠的新生肌肉中快肌纤维数量仅约为野生型小鼠肌肉的2/3。因此,Bcl-2功能在胎儿期和出生后肌生成的特定阶段是必需的。