Sprouty proteins role in division and differentiation of forebrain progenitors

描述（由申请方提供）：必须精细控制受体酪氨酸激酶（RTK）信号传导，以确保正常发育。最近，RTK途径中的一种新的参与者已经被描述-Sprouty（Spry）蛋白。这些蛋白质微调生长因子信号传导，因此可以在神经祖细胞内介导不同的RTK依赖性结果中发挥作用。我们的长期目标是了解Spry蛋白在CNS干细胞中的功能。为了解决这个问题，我们将描述Spry 1和2在发育和成人大脑中的表达模式，并将使用体内和体外技术来阐明其功能。目的1将是表征Sprouty 1和2在胚胎皮层和成人SVZ中的表达。为了揭示Spry 1和Spry 2在前脑生发区的表达，我们将研究mRNA和蛋白质的表达在小鼠大脑皮层发育过程中，并在成人SVZ使用原位杂交和免疫组织化学。在目标2中，我们将确定Sprouty 1和Sprouty 2在胚胎大脑皮层和成人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，我们将使用子宫内电穿孔将Spry 1和2 shRNA或过表达构建体递送到胚胎生发区。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人SVZ培养物中表达Spry 1和2 shRNA或过表达构建体。这些分析，加上表达分析，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人SVZ的功能。神经祖细胞（NPC），包括神经干细胞（NSC），具有许多治疗应用的潜力，包括它们在神经修复中的直接用途和具有干/祖细胞组分的疾病建模的可能性。RTK信号传导在整个发育过程中严重影响干细胞行为，并且RTK信号传导的改变在CNS肿瘤中普遍存在。因此，重要的是要了解RTK信号，以了解正常干细胞生物学和脑癌细胞的关键方面，这可能是从干细胞样细胞产生的。

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.