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A Study on the Role of Arylsulfatase in Sea Urchin Development

芳基硫酸酯酶在海胆发育中的作用研究

基本信息

批准号：
01540591
负责人：
YAMADA Kazumi
金额：
$ 1.22万
依托单位：
Hiroshima University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1989
资助国家：
日本
起止时间：
1989 至 1990
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-01540591/
关键词：
Sea urchin Development Arylsulfatase Gene cloning In situ hybridization Activity staining DNA synthesis Gastrulation ウニ胚活性染色分子発生学

项目摘要

In development of sea urchin embryos, the arylsulfatase activity begins to make a drastic increase at the mesenchyme blastula stage prior to the onset of gastrulation. The present research project has been undertaken to elucidate roles of arylsulfatase in the process of gastrulation. The following are the result of the two-years research project.(1) The arylsulfatase (Ars) gene of the embryo of the sea urchin, Hemicentrotus pulcherrimus, was cloned and its sequence was determined.(2) By in situ hybridization using anti-sense RNA of Ars-cDNA as the probe, it was found that the Ars gene is transcribed only in aboral ectoderm of sea urchin gastrula.(3) By the activity staining of arylsulfatase with optical as well as electron microscopes, it was found that arylulfatase localizes in lysosomes of qastrula, prism and pluteus larvae. It is suggested that arylsulfatase supplies sulfate ions through hydrolysis of yolk material which are needed to support gastrulation.(4) Addition of aphidicolin, an inhibitor of DNA synthesis, to sea urchin embryos resulted in the prevention of transcription of the Ars gene. The result was interpreted as that timing of the Ars gene transcription is somehow related to cycles of DNA replication after fertilization.

在海胆胚胎的发育过程中，芳基硫酸酯酶活性在原肠胚形成开始之前的间充质囊胚阶段开始急剧增加。本研究旨在阐明芳基硫酸酯酶在原肠胚形成过程中的作用。以下是两年研究项目的结果。(1)克隆了马粪海胆（Hemicentrotus pulcherrimus）胚胎的芳基硫酸酯酶（Ars）基因，并测定了其序列。(2)以Ars基因cDNA的反义RNA为探针进行原位杂交，发现Ars基因仅在海胆原肠胚的离口外胚层中转录。(3)通过对芳基硫酸酯酶活性的光学显微镜和电子显微镜染色，发现芳基硫酸酯酶主要定位于扁管虫、棱柱虫和青虫幼虫的溶酶体中。这表明芳基硫酸酯酶通过水解支持原肠胚形成所需的卵黄物质来提供硫酸根离子。(4)此外，阿非迪霉素，DNA合成的抑制剂，海胆胚胎导致的Ars基因的转录的预防。这一结果被解释为Ars基因转录的时间在某种程度上与受精后的DNA复制周期有关。