Direct cell lineage analysis in Drosophila melanogaster by time-lapse, three-dimensional optical microscopy of living embryos.
Direct cell lineage analysis in Drosophila melanogaster by time-lapse, three-dimensional optical microscopy of living embryos.
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DOI：
10.1083/jcb.109.2.505
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发表时间：
1989-08
影响因子：
7.8
通讯作者：
Alberts, B M
中科院分区：
文献类型：
--
作者：
Minden, J S;Agard, D A;Sedat, J W;Alberts, B M
果蝇胚胎细胞分化的最初迹象之一发生在受精后 3 小时，此时离散的细胞组以空间和时间模式方式进入第十四次有丝分裂，形成有丝分裂域（Foe，V.E.和 G.M.Odell，1989，Am.Zool.29：617-652）。为了确定有丝分裂域中的细胞驻留是否仅由早期胚胎中的细胞位置决定，或者细胞谱系是否也发挥作用，我们开发了一种直接分析活胚胎中细胞核行为的技术。通过将荧光标记的组蛋白显微注射到合胞体胚胎中，使用配备高分辨率电荷耦合装置的显微镜记录每个核的运动和分裂，而不会干扰发育。从三维延时记录中生成了两种类型的发育图：一种是在胚胎的一个小区域中追踪每个核从核周期 11 到核周期 14 的谱系历史；另一个根据第14核分裂的时间和模式记录核命运。通过比较两个胚胎的谱系和命运图，我们得出结论，至少对于检查区域，果蝇有丝分裂域形成的模式是由每个细胞的位置决定的，不受细胞谱系的影响。
One of the first signs of cell differentiation in the Drosophila melanogaster embryo occurs 3 h after fertilization, when discrete groups of cells enter their fourteenth mitosis in a spatially and temporally patterned manner creating mitotic domains (Foe, V. E. and G. M. Odell, 1989, Am. Zool. 29:617-652). To determine whether cell residency in a mitotic domain is determined solely by cell position in this early embryo, or whether cell lineage also has a role, we have developed a technique for directly analyzing the behavior of nuclei in living embryos. By microinjecting fluorescently labeled histones into the syncytial embryo, the movements and divisions of each nucleus were recorded without perturbing development by using a microscope equipped with a high resolution, charge-coupled device. Two types of developmental maps were generated from three-dimensional time-lapse recordings: one traced the lineage history of each nucleus from nuclear cycle 11 through nuclear cycle 14 in a small region of the embryo; the other recorded nuclear fate according to the timing and pattern of the 14th nuclear division. By comparing these lineage and fate maps for two embryos, we conclude that, at least for the examined area, the pattern of mitotic domain formation in Drosophila is determined by the position of each cell, with no effect of cell lineage.