Dyce Paul W, Li Dan, Barr Kevin J, Kidder Gerald M
1 Department of Physiology and Pharmacology, Schulich School of Medicine and Dentistry, The University of Western Ontario , London, Ontario, Canada .
Stem Cells Dev. 2014 Jul 15;23(14):1636-46. doi: 10.1089/scd.2013.0459. Epub 2014 May 15.
Expression of the gap junction protein, connexin43 (Cx43), begins early during embryogenesis and is maintained in many different cell types. Several stem cell populations have been shown to express Cx43 and to form functional gap junctions. While it is clear that Cx43 is critical to the function of many organs, whether the same is true for stem cells has not been clearly demonstrated. Recently, stem cells isolated from newborn mouse skin were shown to form oocyte-like cells (OLCs) in vitro, hence the present study focussed on the role Cx43 plays in the proliferation and differentiation of these cells. The stem cells express Cx43 and those from knockout mice (Cx43 KO) exhibited significantly reduced cell-cell coupling. Loss of Cx43 reduced the rate of cellular migration [Cx43 KO, 1.57±0.65 radial cell units (RCU); wildtype (WT), 5.57±0.37 RCU] but increased the proliferation rate of the stem cells (Cx43 KO, 29.40%±2.02%; WT, 12.76%±1.50%). The expression of the pluripotency markers OCT4 and Nanog were found to be reduced in the Cx43 KO population, suggesting an inhibition of differentiation potential. To test the differentiation ability, the stem cells were induced to form neuronal cell types in vitro. While both the WT and KO cells were able to form GFAP-positive astrocytic cells, only WT stem cells were able to form βIII tubulin-positive neurons. Similarly, the ability of the stem cells to form OLCs was ablated by the loss of Cx43. These data reveal a role for Cx43 in maintaining multipotency within the skin-derived stem cell population.
缝隙连接蛋白连接蛋白43(Cx43)在胚胎发育早期就开始表达,并在许多不同细胞类型中持续存在。已有研究表明,几种干细胞群体可表达Cx43并形成功能性缝隙连接。虽然很明显Cx43对许多器官的功能至关重要,但对于干细胞而言是否如此尚未得到明确证实。最近,从新生小鼠皮肤分离的干细胞在体外被证明可形成卵母细胞样细胞(OLC),因此本研究聚焦于Cx43在这些细胞增殖和分化中所起的作用。这些干细胞表达Cx43,而来自基因敲除小鼠(Cx43 KO)的干细胞表现出明显降低的细胞间偶联。Cx43的缺失降低了细胞迁移速率[Cx43 KO,1.57±0.65个径向细胞单位(RCU);野生型(WT),5.57±0.37 RCU],但提高了干细胞的增殖速率(Cx43 KO,29.40%±2.02%;WT,12.76%±1.50%)。发现多能性标志物OCT4和Nanog在Cx43 KO群体中的表达降低,表明分化潜能受到抑制。为了测试分化能力,将这些干细胞在体外诱导形成神经元细胞类型。虽然WT和KO细胞都能够形成GFAP阳性的星形胶质细胞,但只有WT干细胞能够形成βIII微管蛋白阳性的神经元。同样,Cx43的缺失消除了干细胞形成OLC的能力。这些数据揭示了Cx43在维持皮肤来源干细胞群体的多能性方面的作用。