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Gene Analysis of Exogastrula-Inducing Peptides in Sea Urchin Embryo

海胆胚胎中外原肠胚诱导肽的基因分析

基本信息

批准号：
63480019
负责人：
ISHIHARA Katsutoshi
金额：
$ 4.16万
依托单位：
Saitama University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (B)
财政年份：
1988
资助国家：
日本
起止时间：
1988 至 1990
项目状态：
已结题

项目摘要

We found some intrinsic factors in mesencyme blastulae of Anthocidaris crassispina which induce exogastrulation when added outside the embryos. This factors were named exogastrula-inducing peptides (EGIPs). We have investigated the physiological actions, the chemical structure and the mechanisms of of gene expression of EGIPs in the embryos.1. Physilolgical actions of EGIP : We studies the effects on cytoskeletones, cell proliferation and extracellular matrix of embryos. Cytoskeletones were visualized by fluorecent stainings with antibodies for tubulin and actin. It was suggested that the arrangements of microtubules and microfilaments in vegital plates were affected by the presence of EGIP. On the other hand, macromeres, mesomeres and were separate in 16-cell stage and the effects of EGIP on the proliferation of each blastomeres were observed. It was found that the cell divisions of only macromeres were temporarily inhibited and restored on onset of exogastrulation.2. Primary structures of EGIP : 4 Kinds of EGIP were isolated which have some similarities in chemical structures. They are simple peptides which are composed of 53-58 amino acid residues and 6 cysteine residues in common. The amino acid sequences of 4 EGIPs were all determined and the position of disulfide bonds were also settled.3. Synthesis of cDNA and the timing of gene expression : Using EGIP-D, we obtained cDNA with 1935 bp near almost total length of genomic DNA. As a result of translation of the cDNA, it revealed that 4 EGIPs were synthesized as a huge molecule of a precursor protein and expressed in gastrula stage of empryo development.

我们在Anthocidaris crassispina 的间质囊胚中发现了一些内在因子，当添加到胚胎外部时，这些内在因子会诱导外原肠胚形成。该因子被命名为外原肠胚诱导肽（EGIP）。我们对EGIPs在胚胎中的生理作用、化学结构及其基因表达机制进行了研究。 1． EGIP的生理作用：我们研究对胚胎细胞骨架、细胞增殖和细胞外基质的影响。通过微管蛋白和肌动蛋白抗体的荧光染色使细胞骨架可视化。有人提出，植物平板中微管和微丝的排列受到 EGIP 存在的影响。另一方面，在16细胞阶段，大粒、中粒和中粒是分开的，并观察EGIP对每个卵裂球增殖的影响。结果发现，在外原肠胚形成开始时，只有大分子的细胞分裂被暂时抑制和恢复。2． EGIP的一级结构：分离出4种EGIP，它们在化学结构上有一些相似之处。它们是简单的肽，共同由 53-58 个氨基酸残基和 6 个半胱氨酸残基组成。 4个EGIP的氨基酸序列均已确定，二硫键位置也已确定。 3． cDNA的合成和基因表达的时机：使用EGIP-D，我们获得了1935bp的cDNA，几乎接近基因组DNA的全长。 cDNA的翻译结果表明，4个EGIP被合成为前体蛋白的大分子，并在胚胎发育的原肠胚阶段表达。