FUNCTION OF AN ENGRAILED-CLASS PROTEIN IN LEECH

水蛭中一种Egrailed级蛋白质的功能

• 批准号: 2378467
• 负责人: DANIEL H SHAIN
• 金额: $ 2.86万
• 依托单位: UNIVERSITY OF CALIFORNIA BERKELEY
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-02-12 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/2378467
• 关键词: Hirudinea; biochemical evolution; embryogenesis; gene expression; homeobox genes; lasers; microinjections; protein structure function

The long term goal of this research is to understand the process of segment formation in diverse animal phyla. This will contribute toward an understanding of how ancestral developmental processes were modified over the course of evolution, resulting in the dramatic differences that are observed between species. To approach this problem, the function of an engrailed-class protein (ht-en) will be examined in the leech, Helobdella triseflalis. engrailed (en) is a homeobox protein originally identified in Drosophila melanogaster. Homologues of en have since been identified in species throughout the animal kingdom, including human. In all species, en-class proteins appear to function in bCLh pattern formation and in neurogenesis. Consequently, determining the role of an en-class protein in a lower eukaryote (i.e., leech) is likely to contribute toward an understanding of normal development, and also to anomalies associated with abnormal development in higher eukaryotes (i.e., human). The function of ht-en will be examined by manipulating its expression during leech embryogenesis. Specifically, ht-en progenitor cells will be ablated using a 485 nm laser beam; ht-en expression will be abrogated with microinjected antisense oligodeoxynucleotides or ribozymes; and ht-en will be expressed ectopically in selected cell lineages. The effects of these manipulations will be measured against a previously observed fused ganglia phenotype, which results upon elimination of ht-en expressing cells during normal leech development.

这项研究的长期目标是了解 各种动物门的段形成。这将有助于 对祖先发展过程的理解 在演变过程中修改，导致戏剧性 物种之间观察到的差异。接近这一点 问题，界面蛋白（HT-EN）的功能将是 在水ech中检查了Helobdella Triseflalis。 grailed（en）是 Homeobox蛋白最初在果蝇中鉴定出来。 此后在整个物种中都发现了EN的同源物 动物王国，包括人类。在所有物种中，en级蛋白质 似乎在BCLH模式形成和神经发生中起作用。 因此，确定恩类蛋白在较低的蛋白质中的作用 真核生物（即水ech）可能有助于 了解正常发展，也了解相关的异常 在高等真核生物（即人）中发育异常。这 HT-EN的功能将通过操纵其表达来检查 在水ech胚胎发生期间。具体而言，HT-EN祖细胞将 使用485 nm激光束烧杯； ht-en表达将是 用微型注射的反义寡脱氧核苷酸废除或 核酶； HT-EN将在选定的单元格中以异位表示 血统。这些操作的影响将针对 先前观察到的融合神经节表型，导致 在正常的水ech发育过程中消除HT-EN表达细胞。