pHMA, a pH-sensitive GFP reporter for cell engulfment, in Drosophila embryos, tissues, and cells.
pHMA, a pH-sensitive GFP reporter for cell engulfment, in Drosophila embryos, tissues, and cells.
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DOI：
10.1002/dvdy.22180
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发表时间：
2010-02
影响因子：
2.5
通讯作者：
Minden, Jonathan S.
中科院分区：
文献类型：
--
作者：
Fishilevich, Elane;Fitzpatrick, James A. J.;Minden, Jonathan S.
通过吞噬作用吞噬凋亡细胞确保去除不需要的和有缺陷的细胞。我们开发了一种用于细胞吞噬的基因编码标记 pHMA，它由 GFP 的 pH 敏感衍生物 pHluorin 组成，融合到 Moesin 的肌动蛋白结合域。在果蝇胚胎和培养细胞的健康细胞中，pHMA 存在于细胞皮质中。在垂死细胞中，pHMA 失去其皮质定位并报告 pH 值适度下降。在胚胎中，垂死的细胞失去顶端接触，然后向基底移动，最终被邻近的细胞或巨噬细胞吞噬。细胞尸体物质被吞噬后很快被强酸化，并在吞噬细胞中持续数小时。 pHMA 信号的变化与细胞凋亡的增加或减少密切相关。这些数据表明 pHMA 是细胞吞噬的有用报告基因，可用于筛选影响细胞吞噬的突变。
Engulfment of apoptotic cells by phagocytosis ensures the removal of unwanted and defective cells. We developed a genetically encoded marker for cell engulfment, pHMA, which consists of the pH-sensitive derivative of GFP, pHluorin, fused to the actin-binding domain of Moesin. In healthy cells of Drosophila embryos and cultured cells, pHMA resides at the cell cortex. In dying cells, pHMA loses its cortical localization and reports a modest decrease in pH. In embryos, the dying cells lose their apical contacts, then move basally and are ultimately engulfed by neighboring cells or macrophages. The cell corpse material is strongly acidified soon after engulfment and persists in the phagocytic cell for several hours. Changes in the pHMA signal correlate well with increases or decreases in apoptosis. These data show that pHMA is a useful reporter for cell engulfment and can be used in screening for mutations that affect cell engulfment.
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