Search for bioactive substances acting on insect neuropeptide receptors and application of them to insect growth requlators.

寻找作用于昆虫神经肽受体的生物活性物质及其在昆虫生长调节剂中的应用。

基本信息

批准号：
08556017
负责人：
SAKUDA Shohei
金额：
$ 6.66万
依托单位：
The University of Tokyo
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (A)
财政年份：
1996
资助国家：
日本
起止时间：
1996 至 1998
项目状态：
已结题

项目摘要

The life cycle of insects as metamorphosis and ecdysis is controlled by a variety of neuropeptides. Recent basic reserach on insects neuropeptodes has made it possible to search for their antagonists or agonists, which are possible candidates for safe and specific insect growth regulators. In this study, we have attempted to search for a bioactive substance which acts on a receptor of bombyxin or prothoracicotropic hormone (PTTH).First, screening was performed with a bioassay system using BM-N4 cells among metabolites of microorganisms. Since a characteristic morphological change of the cells was induced by bombyxin, an agonistic or antagonistic activity toward bombyxin could be detected by the assay. Unfortunately, no specific bioactive substance on bombyxin receptor was found this time. However, we isolated nocardamine as an inducer of a morphological change of the cells and found a new acidic polysaccharide from Gordona sp. which strongly induced cell aggregation. Next, anew bioassay method with an optical biosensor system was developed to observe an interaction between bonbyxin and BM-N4 cells directly. Since bombyxin clearly inhibited the biding of immobilized biotinylated bombyxin and the cells, we are performing screening bombyxin antagonists using the assay system.As amino acid sequence of PTTH receptor has not been characterized, an expression cloning of the receptor was performed to identify the cDNA encoding the receptor and to construct transgenic COS 7 cells with PTTH receptor. In spite of many challenges, we have not succeeded the cloning yet. From now on, based on the results obtained by this study, we will continue screening search for bioactive substances which act on insect neuropeptide receptors.

昆虫作为变形和肌肉的生命周期受多种神经肽的控制。最近对昆虫神经肽的基本复兴使他们可以寻找其拮抗剂或激动剂，这可能是安全和特定的昆虫生长调节剂的候选者。在这项研究中，我们试图寻找一种生物活性物质，该物质作用于Bombyxin或促肌动型激素（PTTH）。首先，使用BM-N4细胞在微生物的代谢物中使用BM-N4细胞进行筛查。由于Bombyxin诱导细胞的特征形态变化，因此可以通过测定检测到对Bombyxin的激动或拮抗活性。不幸的是，这次没有在Bombyxin受体上发现特定的生物活性物质。但是，我们将甲胺分离为细胞形态变化的诱导剂，并从Gordona SP中发现了新的酸性多糖。强烈诱导细胞聚集。接下来，开发了具有光学生物传感器系统的新生物测定方法，以直接观察Bonbyxin和BM-N4细胞之间的相互作用。由于Bombyxin清楚地抑制了固定生物素化的Bombyxin和细胞的竞标，因此我们使用分析系统进行了筛选Bombyxin拮抗剂。尚未表征PTTH受体的氨基酸序列，对受体的表达进行了表达，以识别cDNA的表达，以识别cdna构造受体和跨基因的细胞。尽管面临许多挑战，但我们还没有成功克隆。从现在开始，根据这项研究的结果，我们将继续筛选搜索对昆虫神经肽受体作用的生物活性物质。