RPG: Preliminary Functional Studies of Hro-twi Protein in the Leech Embryo

RPG：水蛭胚胎中 Hro-twi 蛋白的初步功能研究

• 批准号: 9727760
• 负责人: Julio Soto
• 金额: $ 1.8万
• 依托单位: Eastern New Mexico University Main Campus
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-01-01 至 1999-06-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=9727760&HistoricalAwards=false
• 关键词: RPG; Preliminary; Functional; Studies; Hro

9727760 Soto The main focus of this research is to study the action of genes involved in the process of mesoderm formation and differentiation in the leech embryo. Morphological information, in addition to information obtained from more recent nucleic acid and genetic analysis, has shown that many developmental processes in invertebrate and vertebrate species are very similar, indicating that these processes may be evolutionarily conserved. An example of one of these fundamental processes gastrulation, involves the formation of mesodermal, endodermal and ectodermal embryonic layers. The ectoderm, or outer layer, gives rise to the epithelium and the nervous system. The mesoderm, or middle layer, gives rise to connective tissue, muscles, blood cells, blood vessels, and some organs. The endoderm, or inner layer, gives rise to the digestive tract and the interior lining of some organs. Determining what regulates the formation of these layers has clear implications for the understanding of both normal and abnormal embryonic development. The main objective of this research is to understand the process of gastrulation at both the cellular and molecular level in a simple organism, the leech embryo. Two different approaches will be used to study gastrulation events in the leech. The first will involve a molecular approach where PCR and DNA cloning will be used to identify genes involved in the process of grastrulation. The second approach is the utilization of cell lineage analysis to identify cells involved in the process of gastrulation and therefore affected by the genes identified by the DNA approach. During the period covered by this planning grant, studies will focus on a candidate of mesodermal determination in the leech embryo, Hro-twi (Soto et al., in press). As part of the planning activities, polyclonal antibodies against Hro-twi protein will be purified and protein expression determined. In addition, preliminary functional analysis will be performed, which will determine th e feasibility of antisense/overexpression approaches to study the function of developmentally-important genes in the leech embryo.

小行星9727760 本研究的主要目的是研究水蛭胚胎中胚层形成和分化过程中相关基因的作用。 形态学的信息，以及从最近的核酸和遗传分析中获得的信息，已经表明无脊椎动物和脊椎动物物种中的许多发育过程非常相似，这表明这些过程可能是进化保守的。 这些基本过程之一的一个例子原肠胚形成涉及中胚层、内胚层和外胚层的形成。 外胚层或外层产生上皮和神经系统。 中胚层或中间层产生结缔组织、肌肉、血细胞、血管和一些器官。 内胚层或内层形成消化道和某些器官的内层。 确定是什么调节这些层的形成对于理解正常和异常胚胎发育具有明确的意义。 本研究的主要目的是在细胞和分子水平上了解水蛭胚胎这一简单生物体的原肠胚形成过程。 将使用两种不同的方法来研究水蛭中的原肠胚形成事件。 第一个将涉及一种分子方法，其中PCR和DNA克隆将被用来确定参与grastrulation过程的基因。 第二种方法是利用细胞谱系分析来鉴定参与原肠胚形成过程的细胞，从而受到DNA方法鉴定的基因的影响。 在这项计划拨款所涵盖的期间，研究将集中在水蛭胚胎中中胚层决定的候选者Hro-twi（Soto等人，印刷中）。 作为计划活动的一部分，将纯化抗Hro-twi蛋白的多克隆抗体并测定蛋白表达。 此外，还将进行初步的功能分析，以确定反义/过表达方法研究水蛭胚胎发育重要基因功能的可行性。