Department of Environmental Biochemistry, Kyoto Pharmaceutical University, 5 Nakauchi-cho, Misasagi, Yamashina-ku, Kyoto 607-8414, Japan.
Biochem Biophys Res Commun. 2013 Sep 13;439(1):90-5. doi: 10.1016/j.bbrc.2013.08.022. Epub 2013 Aug 16.
We previously demonstrated that P2X7 receptors (P2X7Rs) expressed by cultured mouse astrocytes were activated without any exogenous stimuli, but its roles in non-stimulated resting astrocytes remained unknown. It has been reported that astrocytes exhibit engulfing activity, and that the basal activity of P2X7Rs regulates the phagocytic activity of macrophages. In this study, therefore, we investigated whether P2X7Rs regulate the engulfing activity of mouse astrocytes. Uptake of non-opsonized beads by resting astrocytes derived from ddY-mouse cortex time-dependently increased, and the uptaken beads were detected in the intracellular space. The bead uptake was inhibited by cytochalasin D (CytD), an F-actin polymerization inhibitor, and agonists and antagonists of P2X7Rs apparently decreased the uptake. Spontaneous YO-PRO-1 uptake by ddY-mouse astrocytes was reduced by the agonists and antagonists of P2X7Rs, but not by CytD. Down-regulation of P2X7Rs using siRNA decreased the bead uptake by ddY-mouse astrocytes. In addition, compared to in the case of ddY-mouse astrocytes, SJL-mouse astrocytes exhibited higher YO-PRO-1 uptake activity, and their bead uptake was significantly greater. These findings suggest that resting astrocytes exhibit engulfing activity and that the activity is regulated, at least in part, by their P2X7Rs.
我们之前已经证实,培养的小鼠星形胶质细胞表达的 P2X7 受体(P2X7Rs)在没有任何外源性刺激的情况下被激活,但它在非刺激的静息星形胶质细胞中的作用尚不清楚。有报道称星形胶质细胞具有吞噬作用,而 P2X7Rs 的基础活性调节巨噬细胞的吞噬活性。因此,在这项研究中,我们研究了 P2X7Rs 是否调节小鼠星形胶质细胞的吞噬作用。非调理珠被来自 ddY 小鼠皮质的静息星形胶质细胞摄取,呈时间依赖性增加,并且摄取的珠被检测到在细胞内空间。细胞松弛素 D(CytD),一种 F-肌动蛋白聚合抑制剂,可抑制珠的摄取,并且 P2X7Rs 的激动剂和拮抗剂明显减少了摄取。ddY 小鼠星形胶质细胞中自发的 YO-PRO-1 摄取被 P2X7Rs 的激动剂和拮抗剂减少,但不受 CytD 影响。使用 siRNA 下调 P2X7Rs 可减少 ddY 小鼠星形胶质细胞的珠摄取。此外,与 ddY 小鼠星形胶质细胞相比,SJL 小鼠星形胶质细胞表现出更高的 YO-PRO-1 摄取活性,并且它们的珠摄取明显更高。这些发现表明,静息星形胶质细胞表现出吞噬作用,并且该活性至少部分受到其 P2X7Rs 的调节。