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Presence and Importance of Connexin43 During Myogenesis

机译:连接蛋白43在肌发生过程中的存在和重要性。

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We analyzed the expression of connexin(Cx)43 in proliferating and differentiating C_2C_(12) cells and in myoblasts obtained from newborn mice. Cx43 was present in both cell types and under both conditions. The functional role of gap junctional communication (GJC) during terminal differentiation was evaluated in C_2C_(12) myoblasts in the presence or absence of the gap junction blocker 18β-glycyrrhetinic acid (β-GA). Differentiation was temporally analyzed through myogenin expression, activity of creatine kinase (CK), and yield of multinucleated cells. In cells treated with β-GA, the CK activity and myotube formation were reversibly blocked. While in control cultures positive myogenin expression was seen in cell clusters, in β-GA treated cultures the myogenin immunoreactivity was detected in few, preferentially sparse cells. The role of Cx43 during terminal differentiation was evaluated in cultures of myoblasts obtained from Cx43~(Cre~ER(T)/fl) transgenic mice. Inducible deletion of Cx43 was obtained upon activation of Cre-ER(T) via 4-OH-tamoxifen applications. Cx43 deletion led to a drastic decrease in myogenin expression at 24 h of differentiation as compared to myoblasts from control mice. Our results indicate that Cx43-containing gap junctions are required for normal skeletal muscle terminal differentiation. These channels might provide a pathway for the intercellular transfer of signals involved in myogenesis.
机译:我们分析了连接蛋白(Cx)43在增殖和分化C_2C_(12)细胞和新生小鼠成肌细胞中的表达。 Cx43存在于两种细胞类型和两种条件下。在存在或不存在间隙连接阻滞剂18β-甘草次酸(β-GA)的情况下,在C_2C_(12)成肌细胞中评估了间隙连接通讯(GJC)在终末分化过程中的功能作用。通过肌生成素的表达,肌酸激酶(CK)的活性和多核细胞的产量来临时分析分化。在用β-GA处理的细胞中,CK活性和肌管形成被可逆地阻断。虽然在对照培养物中,在细胞簇中见到阳性的肌细胞生成素表达,但在β-GA处理的培养物中,在少数稀疏的细胞中检测到了肌生成素免疫反应性。在从Cx43〜(Cre〜ER(T)/ fl)转基因小鼠获得的成肌细胞培养物中,评估了Cx43在终末分化中的作用。通过4-OH-他莫昔芬应用激活Cre-ER(T)后,获得了Cx43的诱导型缺失。与来自对照小鼠的成肌细胞相比,Cx43缺失导致分化后24 h的肌生成素表达急剧下降。我们的结果表明,正常骨骼肌末端分化需要Cx43间隙连接。这些通道可能为参与肌发生的信号的细胞间转移提供途径。

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