PROGRAMMED CELL-DEATH IN THE INTERDIGITAL TISSUE OF THE FETAL MOUSE LIMB IS APOPTOSIS WITH DNA FRAGMENTATION
PROGRAMMED CELL-DEATH IN THE INTERDIGITAL TISSUE OF THE FETAL MOUSE LIMB IS APOPTOSIS WITH DNA FRAGMENTATION
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DOI：
10.1002/ar.1092420114
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发表时间：
1995-05-01
影响因子：
--
通讯作者：
SHIOTA, K
中科院分区：
文献类型：
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作者：
MORI, C;NAKAMURA, N;SHIOTA, K
背景:程序性细胞死亡是哺乳动物形态发生过程中必不可少的事件，它消除了不需要的细胞来完成组织发生和器官发生，发育中的肢体指间间隙的细胞死亡是形态发生细胞死亡的典型例子。我们研究了小鼠发育肢体指间组织的典型程序性细胞死亡是否是核DNA断裂的细胞凋亡，并对小鼠胎儿发育肢体中细胞程序性死亡和细胞增殖的发生进行了序贯研究，分析了它们之间的相互关系。方法：采用硫酸尼罗蓝染色、琼脂糖凝胶电泳检测DNA阶梯、细胞化学标记DNA片段等方法检测小鼠胎儿发育肢体中凋亡细胞死亡的发生情况。我们还使用BrdU/抗BrdU免疫组织化学标记增殖细胞，并检测凋亡程序性细胞死亡与细胞增殖之间的相互关系。结果：琼脂糖凝胶电泳法在13天小鼠胎指间组织DNA提取液中检测到细胞凋亡的生化证据DNA阶梯。通过尼罗蓝染色和DNA片段的细胞化学标记分别在指间中皮层和指关节区域检测到程序性细胞死亡和DNA片段，用于鉴定增殖s期细胞的BrdU/抗BrdU免疫组织化学显示，指间间质细胞在程序性细胞死亡和DNA片段开始之前停止DNA合成。结论：我们证实了胚胎小鼠四肢指间组织和推定关节区发生细胞学凋亡改变和核DNA断裂，它们似乎在指间分离和关节腔的形成中起重要作用。(C) 1995 Wiley-Liss, Inc。
Background: Programmed cell death is an essential event during mammalian morphogenesis which eliminates unnecessary cells to accomplish histogenesis and organogenesis, Cell death in interdigital spaces of the developing limb is a classical example of morphogenetic cell death, We investigated whether classical programmed cell death in the interdigital tissue of the developing limb in mice is apoptosis with fragmentation of nuclear DNA and also examined sequentially the occurrence of programmed cell death and cell proliferation in the developing limb of mouse fetuses to analyze their interrelation.Methods: We examined the occurrence of apoptotic cell death in the developing limbs of mouse fetuses by using Nile blue sulphate staining, agarose gel electrophoresis for detecting DNA laddering, and a cytochemical labeling of DNA fragmentation. We also labeled proliferating cells using BrdU/anti-BrdU immunohistochemistry and examined the interrelation between apoptotic programmed cell death and cell proliferation.Results: DNA ladders, a biochemical evidence of apoptosis, were detected in DNA extracts from the interdigital tissue of day 13 mouse fetuses by agarose gel electrophoresis. Programmed cell death and DNA fragmentation were detected by Nile blue staining and cytochemical labeling of DNA fragmentation, respectively, in the interdigital mesoderm and in the regions of presumptive joints of the digit, BrdU/anti-BrdU immunohistochemistry for identifying proliferating S-phase cells revealed that interdigital mesenchymal cells cease DNA synthesis before programmed cell death and DNA fragmentation begin.Conclusions: We confirmed that both cytological apoptotic alterations and fragmentation of nuclear DNA occur in the interdigital tissue and presumptive joint areas of fetal mouse limbs, and they appear to play a significant role in the separation of digits as well as the formation of joint cavities. (C) 1995 Wiley-Liss, Inc.