NOTCH GENES IN DEVELOPMENT

描述（改编自申请人摘要）：Weinmaster博士 目前的假设是，像果蝇Notch这样的基因， 有助于确定细胞命运的细胞间相互作用的子集。 的 具体目标旨在提供所需的信息， Notch基因在哺乳动物发育过程中的作用。 使用 原位杂交和免疫组织化学技术，她将 确定细胞类型和发育过程， 这两个密切相关的基因被表达。 提供的资料 这些研究将确定候选细胞间相互作用介导的 通过Notch基因，这可能被用于细胞培养， 测量Notch活性的系统。 关于Notch是否 介导细胞间相互作用将使用神经元-雪旺氏 细胞共培养。 如果Notch介导了神经元和 施旺细胞然后基因工程Notch-减去施旺细胞 将无法对神经元接触做出反应。 本构 Notch在转染的C2 C12成肌细胞中的表达阻止了肌管 形成，提供方便的Notch活性“读出”。 博士 Weinmaster将使用这种分析来确定 抑制C2 C12肌源性分化所需的Notch 从而开始研究Notch的机制 功能 这些细胞培养实验的重要补充 就是分离Notch的配体 细胞表达 然后可以将Notch或其配体共培养，并将下游Notch或其配体与细胞共培养。 通过Notch介导的细胞间相互作用的影响， 监测以确定陷波信号所需的事件 转导 这项工作的最终目标是定义 Notch 1和Notch 2在哺乳动物细胞中的功能和作用方式 在正常的发展过程中。

DESCRIPTION (adapted from applicant's abstract): Dr. Weinmaster's working hypothesis is that genes like Drosophila Notch, mediate a subset of cell-cell interactions that help specify cell fate. The specific aims are designed to provide information required to reveal the role that Notch genes play during mammalian development. Using in situ hybridization and immunohistochemical techniques she will determine the cell types and the developmental processes in which these two closely related genes are expressed. Information provided by these studies will identify candidate cell-cell interactions mediated by the Notch genes, which could potentially be used as cell culture systems to measure Notch activity. The question of whether Notch mediates cell-cell interactions will be studied using neuron-Schwann cell cocultures. If Notch mediates interactions between neurons and Schwann cells then genetically engineered Notch-minus Schwann cells will be unable to respond to neuronal contact. Constitutive expression of Notch in transfected C2C12 myoblasts prevents myotube formation, providing a convenient "read-out" of Notch activity. Dr. Weinmaster will use this assay to determine the structural motifs of Notch that are required to inhibit myogenic differentiation of C2C12 cells and thereby begin to investigate the mechanisms of Notch function. An important complement to these cell culture experiments would be the isolation of ligands for Notch. Cells expressing either Notch or its ligand could then be cocultured and the downstream effects of cell-cell interactions, mediated through Notch, would be monitored to determine the events required for Notch- signal transduction. The ultimate goal of this work is to define the function and mode of action of Notch1 and Notch2 in mammalian cells during normal development.