Salas-Vidal E, Lomelí H, Castro-Obregón S, Cuervo R, Escalante-Alcalde D, Covarrubias L
Departmento de Genética y Fisiología Molecular, Universidad Nacional Autónoma de México, Cuernavaca, Morelos, México.
Exp Cell Res. 1998 Jan 10;238(1):136-47. doi: 10.1006/excr.1997.3828.
Programmed cell death or apoptosis is an essential process during the morphogenesis of a large number of structures. Evidence obtained over the past few years indicates that, in some cases, the generation of reactive oxygen species (ROS) is an important event during the course of apoptosis. Using an in vitro culture system in which digit individualization of developing limbs normally occurs, we assayed the effect of different antioxidants on the cell death that takes place at interdigits. The addition of phenol, dimethyl sulfoxide, or 2',7'-dichlorodihydrofluorescein diacetate (DCDHF-DA) to murine developing limbs in culture prevented digit individualization as well as the typical interdigital cell death. Two ROS-sensitive dyes, 3-(4,5-dimethylthiazol)-2,5-diphenyl tetrazolium bromide and DCDHF-DA, stained interdigits and the so-called "necrotic zones," implying that they contain cells under oxidative stress. Very few interdigital cells were doubly stained with the ROS probes and two cell death indicators (i.e., acridine orange and propidium iodide), suggesting that they detect a different stage during the course of apoptosis. Furthermore, we found cells stained for ROS that did not express a specific macrophage marker and in a few cases were seen surrounded by a macrophage. Surprisingly, many regions of the midgestation mouse embryo that are undergoing cell death correlated with those that have a markedly higher level of ROS. Our data suggest that the generation of oxidative stress is a common requirement for cell death that occurs during mouse embryonic development.
程序性细胞死亡或凋亡是许多结构形态发生过程中的一个重要过程。过去几年获得的证据表明，在某些情况下，活性氧（ROS）的产生是凋亡过程中的一个重要事件。我们使用一种体外培养系统，在该系统中发育中的肢体通常会发生指（趾）分化，检测了不同抗氧化剂对指间发生的细胞死亡的影响。向培养中的小鼠发育肢体中添加苯酚、二甲基亚砜或2',7'-二氯二氢荧光素二乙酸酯（DCDHF-DA）可阻止指（趾）分化以及典型的指间细胞死亡。两种对ROS敏感的染料，3-（4,5-二甲基噻唑）-2,5-二苯基溴化四氮唑和DCDHF-DA，对指间和所谓的“坏死区”进行了染色，这意味着它们含有处于氧化应激状态的细胞。很少有指间细胞同时被ROS探针和两种细胞死亡指示剂（即吖啶橙和碘化丙啶）双重染色，这表明它们检测的是凋亡过程中的不同阶段。此外，我们发现被ROS染色的细胞不表达特定的巨噬细胞标志物，并且在少数情况下可以看到它们被巨噬细胞包围。令人惊讶的是，妊娠中期小鼠胚胎中许多正在发生细胞死亡的区域与那些ROS水平明显较高的区域相关。我们的数据表明，氧化应激的产生是小鼠胚胎发育过程中发生细胞死亡的一个共同条件。
