Sprouty proteins role in division and differentiation of forebrain progenitors

描述(申请人提供)：受体酪氨酸激酶(RTK)信号必须被精细地控制以确保正常发育。最近，RTK途径中的一个新的参与者被描述-Sprouty(Spry)蛋白。这些蛋白质微调生长因子信号，因此可能在神经前体细胞内调节不同的RTK依赖结果中发挥作用。我们的长期目标是了解Spry蛋白在中枢神经系统干细胞中的功能。为了解决这个问题，我们将描述Spry1和Spry2在发育中和成年脑中的表达模式，并将使用体内和体外技术来阐明它们的功能。目的1研究Sprouty 1和Sprouty 2在胚胎皮质和成体SVZ中的表达。为了揭示Spry1和SPRY2在前脑生发区的表达，我们将利用原位杂交和免疫组织化学方法检测发育过程中小鼠大脑皮层和成年SVZ中的mRNA和蛋白表达。在目标2中，我们将确定Sprouty1和Sprouty2在胚胎大脑皮层和成年SVZ中的功能。为此，我们将使用体内和体外技术。对于体内分析，我们将使用宫内电穿孔将Spry1和2shRNA或过表达构建物传递到胚胎生发区。对于体外试验，我们将使用慢病毒递送系统在胚胎大脑皮层和成年SVZ培养中表达Spry1和2shRNA或过表达构建物。这些分析结合表达分析，将使我们能够确定这些新的RTK抑制剂在发育中的胚胎大脑皮层和成年SVZ中的功能。神经前体细胞(NPC)，包括神经干细胞(NSCs)，具有许多治疗应用的潜力，包括它们在神经修复中的直接使用，以及对含有干细胞/祖细胞成分的疾病进行建模的可能性。RTK信号在整个发育过程中严重影响干细胞的行为，RTK信号的改变在中枢神经系统肿瘤中普遍存在。因此，为了了解正常干细胞生物学和脑癌细胞的关键方面，了解RTK信号转导是很重要的，这些细胞可能来自干细胞样细胞。

DESCRIPTION (provided by applicant): Receptor Tyrosine Kinase (RTK) signaling must be delicately controlled to ensure normal development. Recently, a novel player in the RTK pathway has been described - the Sprouty (Spry) proteins. These proteins fine-tune growth factor signaling, and could thus play a role in mediating different RTK-dependent outcomes within neural progenitor cells. Our long term goal is to understand Spry protein function in CNS stem cells. To address this we will describe the expression patterns of Spry1 and 2 in the developing and adult brain, and will use both in vivo and in vitro techniques to elucidate their function. Aim 1 will be to characterize the expression of Sprouty 1 and 2 in the embryonic cortex and the adult SVZ. To reveal Spry1 and Spry2 expression in forebrain germinal zones, we will examine mRNA and protein expression in the mouse cerebral cortex during development and in the adult SVZ using in situ hybridization and immunohistochemistry. In aim 2 we will determine the function of Sprouty1 and Sprouty2 in the embryonic cerebral cortex and adult SVZ. For this aim we will use both in vivo and in vitro techniques. For in vivo analysis we will use in utero electroporation to deliver Spry1 and 2 shRNA or overexpression constructs to the embryonic germinal zone. For in vitro assays, we will use a lentiviral delivery system to express Spry1 and 2 shRNA or overexpression constructs in embryonic cerebral cortical and adult SVZ cultures. These assays, coupled with the expression analysis, will allow us to determine the function of these novel RTK inhibitors in the developing embryonic cerebral cortex and adult SVZ. Neural progenitor cells (NPCs), which include neural stem cells (NSCs) have the potential for numerous therapeutic applications, including their direct use in neural repair and the possibility of modeling diseases that have a stem/progenitor cell component. RTK signaling critically impacts stem cell behavior throughout development, and alterations in RTK signaling are prevalent in CNS tumors. Hence it is important to understand RTK signaling in order to understand key aspects of normal stem cell biology and brain cancer cells, which may arise from a stem-like cell.