The Cellular Mechanisms Responsible for AP-2-Dependent Mammalian Morphogenesis

负责 AP-2 依赖性哺乳动物形态发生的细胞机制

• 批准号: 9810504
• 负责人: Trevor Williams
• 金额: $ 22万
• 依托单位: Yale University
• 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: 1998-09-01 至 2000-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=9810504&HistoricalAwards=false
• 关键词: Cellular; Mechanisms; Responsible; AP; Dependent

Sequence-specific DNA binding transcription factors are powerful regulators of genetic information and are responsible for integrating gene expression during embryogenesis. A comprehensive analysis of how these proteins control cell fate determination will yield critical insight into the mechanisms of vertebrate development. The AP-2 protein presents a particularly attractive system to study the various processes governing developmentally-regulated gene expression. During embryogenesis, AP-2 expression is concentrated in many tissues undergoing complex morphogenic changes - the developing neural crest, face, limb bud, kidney, mammary gland and epidermis. In addition, transcription of the AP-2 gene is influenced by retinoic acid, and AP-2 may act as an integral component of this signal transduction pathway.Mice containing a homozygous disruption of the AP-2 gene die perinatally and exhibit multiple and severe developmental defects. Furthermore, preliminary analysis of chimeric embryos, composed of wild-type and AP-2-null cells, indicates that AP-2 is independently required for at least five fundamental morphogenic programs - formation of the neural tube, face, eye, body-wall and limbs. Clearly, a thorough understanding of the cell fate decisions regulated by AP-2 will yield a wealth of information concerning these normal development programs.This proposal will utilize knowledge of the AP-2 knockout phenotype to probe the cellular mechanisms regulating mammalian morphogenesis. A new generation of AP-2-null embryonic stem cells will be established that are tagged with a lacZ transgene. These ES cells will then be used to generate chimeric embryos composed of wild-type and AP-2-null cells. Analysis of these chimeric animals will provide detailed information concerning the cellular basis of the numerous developmental programs with which AP-2 is involved.

序列特异性DNA结合转录因子是遗传信息的强有力的调节因子，并且负责在胚胎发生期间整合基因表达。对这些蛋白质如何控制细胞命运决定的全面分析将对脊椎动物发育的机制产生重要的见解。AP-2蛋白提出了一个特别有吸引力的系统来研究发育调控基因表达的各种过程。在胚胎发生过程中，AP-2表达集中在许多经历复杂形态发生变化的组织中-发育中的神经嵴、面部、肢芽、肾脏、乳腺和表皮。此外，AP-2基因的转录受视黄酸的影响，AP-2可能是这一信号转导通路的组成部分，含有AP-2基因纯合破坏的小鼠在围产期死亡，并表现出多种和严重的发育缺陷。此外，对由野生型和AP-2缺失细胞组成的嵌合体胚胎的初步分析表明，AP-2是至少五个基本形态发生程序-神经管、面部、眼睛、体壁和四肢形成所必需的。显然，对AP-2调控的细胞命运决定的透彻理解将产生关于这些正常发育程序的丰富信息，该提议将利用AP-2敲除表型的知识来探测调节哺乳动物形态发生的细胞机制。将建立用lacZ转基因标记的新一代AP-2缺失胚胎干细胞。然后，这些ES细胞将用于产生由野生型和AP-2缺失细胞组成的嵌合胚胎。对这些嵌合体动物的分析将提供有关AP-2参与的众多发育程序的细胞基础的详细信息。