The signaling role of Qsulf in embryonic neural tube

Qsulf在胚胎神经管中的信号作用

• 批准号: 6626175
• 负责人: Xingbin Ai
• 金额: $ 1.17万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-06-01 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6626175
• 关键词: binding proteins; biological signal transduction; complementary DNA; developmental genetics; developmental neurobiology; electroporation; embryogenesis; enzyme activity; extracellular matrix; gene expression; glucosamine; growth factor; immunocytochemistry; immunoprecipitation; in situ hybridization; neural plate /tube; postdoctoral investigator; proteoglycan; quail; sulfatases; tissue /cell culture; yeast two hybrid system

DESCRIPTION (provided by applicant) The objective of this proposal is to characterize the signaling roles of two candidate heparan sulfate proteoglycan (HSPG) modifying enzymes, quail glucosamine 6-O-sulfatase 1 (QSulf1) and QSulf2 in developing neural tube. Cell surface HSPGs play important roles in modulating the distribution and/or activity of signaling molecules such as Wnts, Shh, FGFs and BMPs. QSulf1, the first known extracellular matrix G6-sulfatase, is required for somite MyoD activation through its enzymatic activity to regulate Wnt pathway. QSulf2 is the second member of the family with unknown function. QSulf1 and QSulf2 have overlapping and discrete expression domains in embryonic neural tube. The proposal will test the hypothesis that QSulf1 and QSulf2 have unique functions in extracellular signaling pathways and the functional distinction is controlled by their divergent hydrophilic domains. First, we will characterize the expression pattern of QSulf1 and QSulf2 during embryogenesis by whole mount in situ hybridization combined with immunohistochemistry. Results from this experiment will serve as the initial steps for the functional analysis of QSulfs in developing neural tube. Second, to define QSulf function in the neural tube, we will manipulate the expression of each gene both by electroporation-mediated overexpression and antisense inhibition in chick embryos at stage 12. Molecular markers for neural tube patterning and neural crest cells will be characterized by immunohistochemistry. Results from these experiments will provide information in signaling roles of QSulfs in embryonic neural tube. Third, co-immunoprecipitation and yeast two-hybrid screen will be conducted to identify the binding proteins for the divergent hydrophilic domains of QSulfs that are hypothesized to control the localized functions of QSuIf1 or QSulf2. This study will provide insights on the mechanisms for localized QSulf function in the embryo.

描述（由申请人提供） 该提议的目的是表征两个的信号传导作用 候选硫酸乙酰肝素蛋白聚糖（HSPG）修饰酶，鹌鹑 葡萄糖胺6-O-硫酸蛋白酶1（QSULF1）和QSULF2在发育的神经管中。细胞 表面HSPG在调节分布和/或 WNT，SHH，FGF和BMP等信号分子的活性。 QSULF1， Somite myod需要首个已知的细胞外基质G6-硫酸酶 通过其酶活性激活以调节Wnt途径。 qsulf2是 功能未知的家庭的第二个成员。 QSULF1和QSULF2具有 胚胎神经管中的重叠和离散表达域。这 建议将检验QSULF1和QSULF2具有独特功能的假设 在细胞外信号通路中，功能区别为 由其发散的亲水结构域控制。首先，我们将描述 QSULF1和QSULF2在胚胎发生过程中的表达模式 原位杂交结合免疫组织化学。结果 实验将作为功能分析的初始步骤 Qsulfs在发展神经管中。第二，在 神经管，我们将通过 电穿孔介导的过表达和反义抑制 第12阶段的胚胎。神经管和神经的分子标记 波峰细胞将以免疫组织化学为特征。这些结果 实验将提供Qsulfs信号作用的信息 神经管。第三，共免疫沉淀和酵母双杂交屏幕将是 进行识别发散亲水性的结合蛋白 Qsulfs的域，该域被认为可以控制 QSUIF1或QSULF2。这项研究将提供有关机制的见解 胚胎中的局部QSULF功能。