Early embryonic development in the spider Achaearanea tepidariorum: Microinjection verifies that cellularization is complete before the blastoderm stage
Early embryonic development in the spider Achaearanea tepidariorum: Microinjection verifies that cellularization is complete before the blastoderm stage
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DOI：
10.1016/j.asd.2010.05.009
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发表时间：
2010-11-01
影响因子：
2
通讯作者：
Oda, Hiroki
中科院分区：
文献类型：
--
作者：
Kanayama, Masaki;Akiyama-Oda, Yasuko;Oda, Hiroki
蜘蛛Achaearanea tepidariorum正在成为研究发育生物学的非昆虫模型。然而，对这种蜘蛛的早期胚胎进行显微注射的可行性尚未见报道。我们确定了白斑拟南芥的早期胚胎阶段，并对其胚胎进行了显微注射。在前胚层16核和32核阶段，能量向卵周移动。当将荧光染料结合的葡聚糖微注射到16核期胚胎的外周区域时，它通常被纳入单个能量并在后代中遗传，而不会泄漏到周围的能量中。这表明16核阶段的胚胎由隔室组成，每个隔室含有一个能量。这些隔室被认为是独立的细胞。荧光染料共轭右旋糖酐可以引入16- 128核期胚胎的单细胞，使我们能够跟踪细胞的命运和运动。注射编码核定位信号/绿色荧光蛋白融合结构的mRNA，证实了该蛋白在活蜘蛛胚胎中的外源表达。我们建议使用显微注射将促进蜘蛛发育的研究。此外，这些数据表明，与果蝇合胞囊胚胚胎不同的是，原胞胚胚的细胞结构限制了细胞质基因产物的扩散。(C) 2010 Elsevier Ltd.版权所有。
The spider Achaearanea tepidariorum is emerging as a non-insect model for studying developmental biology. However, the availability of microinjection into early embryos of this spider has not been reported. We defined the early embryonic stages in A. tepidariorum and applied microinjection to its embryos. During the preblastoderm 16-and 32-nucleus stages, the energids were moving toward the egg periphery. When fluorochrome-conjugated dextran was microinjected into the peripheral region of 16-nucleus stage embryos, it was often incorporated into a single energid and inherited in the progeny without leaking out to surrounding energids. This suggested that 16-nucleus stage embryos consisted of compartments, each containing a single energid. These compartments were considered to be separate cells. Fluorochrome-conjugated dextran could be introduced into single cells of 16-to 128-nucleus stage embryos, allowing us to track cell fate and movement. Injection with mRNA encoding a nuclear localization signal/green fluorescent protein fusion construct demonstrated exogenous expression of the protein in live spider embryos. We propose that use of microinjection will facilitate studies of spider development. Furthermore, these data imply that in contrast to the Drosophila syncytial blastoderm embryo, the cell-based structure of the Achaearanea blastoderm embryo restricts diffusion of cytoplasmic gene products. (C) 2010 Elsevier Ltd. All rights reserved.