Molecular control of intestinal regeneration

肠道再生的分子控制

• 批准号: 7657287
• 负责人: JOSE E GARCIA-ARRARAS
• 金额: $ 35.53万
• 依托单位: UNIVERSITY OF PUERTO RICO RIO PIEDRAS
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-08-01 至 2012-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7657287
• 关键词: Animals; Biological Models; Candidate Disease Gene; Cell physiology; Cells; Cessation of life; Computer Simulation; Databases; Development; Enteric Nervous System; Event; Expressed Sequence Tags; Gastrointestinal tract structure; Genes; Growth; Holothuria; Human; Immunohistochemistry; In Situ Hybridization; Intestines; Laboratories; Lead; Methods; Molecular; Morphology; Natural regeneration; Organ; Pattern; Pharmaceutical Preparations; Process; RNA Interference; Sea Cucumbers; Tissues; Vertebrates; Western Blotting; base; blastema; gene function; knock-down; nerve supply; novel; novel therapeutics; population migration; regenerative; research study; tool

DESCRIPTION (provided by applicant): The capability to regenerate lost or damaged organs is severely limited in humans and most vertebrate animals. Other animals, in contrast, have remarkable regenerating abilities. Among these are the echinoderms, a group of animals closely related to vertebrates. We have been using the sea cucumber Holothuria glaberrima as a model system to study intestinal regeneration. These animals can regenerate their complete digestive tract following a process of auto-evisceration. Our laboratory has characterized the intestinal regeneration process and the associated cellular events. In recent years we have begun to identify genes that might control different aspects of the regeneration process by using molecular tools, particularly the construction and analysis of an EST database and the use of microarrays. These experiments have led to the identification of candidate genes differentially expressed during intestinal regeneration. The present proposal focuses on 20 of these genes. We propose to use these candidate genes to study the molecular control of intestinal regeneration. We intend to validate their differential expression and characterize their sequence, spatial and temporal expression patterns, using in silico methods as well as QRT-PCR, Western blotting, in situ hybridization and immunohistochemistry. This will lead to determining the cellular or tissue component where they are being expressed and a putative function for the genes. More importantly, we will determine the gene's function by knock-down experiments using interference RNA (RNAi) in conjunction with various markers that serve to determine the cellular processes that occur during regeneration such as: changes in tissue cellular components and morphology, formation of the regeneration blastema, intestinal lumen, enteric nervous system, and tissue innervation, as well as changes in cell populations, migration, division, death, dedifferentiation, and growth among others. Our results will help discover novel genes associated with intestinal regeneration and characterize their expression to particular cells and regenerative events. Moreover, their function will be determined, and mechanisms of how they might control the intestinal regeneration process will be identified. We expect that our experiments will make available information, that can be used to study the limitations of vertebrate regenerative capacities and that will serve as the basis for the development of new therapeutics/drugs that modulate regenerative processes.

描述（由申请人提供）：在人和大多数脊椎动物中，再生损失或受损的器官的能力受到严重限制。相比之下，其他动物具有显着的再生能力。其中包括棘皮动物，一组与脊椎动物密切相关的动物。我们一直在使用海参全肠glaberrima作为研究肠道再生的模型系统。这些动物可以在自动visc虫过程后再生其完全消化道。我们的实验室表征了肠再生过程和相关的细胞事件。近年来，我们已经开始通过使用分子工具，尤其是对EST数据库的构建和分析和微阵列的使用来确定可能控制再生过程不同方面的基因。这些实验导致鉴定在肠再生过程中差异表达的候选基因。目前的提案重点介绍了这些基因的20个。我们建议使用这些候选基因研究肠再生的分子控制。我们打算验证它们的差异表达，并使用硅方法以及QRT-PCR，蛋白质印迹，原位杂交和免疫组织化学来表征它们的序列，空间和时间表达模式。这将导致确定它们表达的细胞或组织成分，并确定基因的推定功能。更重要的是，我们将通过使用干扰RNA（RNAI）结合各种标记来确定基因的功能，以确定重新生成过程中发生的细胞过程，例如：组织细胞成分的变化和形成的变化，形成的胚胎，肠道，肠道，肠道神经系统和组织，以及细胞的繁殖，以及细胞，以及细胞的繁殖，以及层次，以及细胞，以及细胞，以及细胞，以及细胞的群体，以及细胞内部的变化，以及范围内部，以及范围内部的变化。去分化和增长。我们的结果将有助于发现与肠道再生有关的新基因，并表征它们对特定细胞和再生事件的表达。此外，将确定它们的功能，并将确定如何控制肠道再生过程的机制。我们预计我们的实验将提供可用的信息，这些信息可用于研究脊椎动物再生能力的局限性，并将作为开发调节再生过程的新疗法/药物的基础。