SSA - The role of neural specific Src splice variants in neuronal specification and differentiation.

(N1-Src和N2-Src)在神经组织中丰富，N-Src的高水平表达与神经母细胞瘤的良好预后有关，导致肿瘤的自发分化和消退。这些观察结果表明，N-SRCS对神经元分化有调节作用。我们以前的工作表明，N1-Src的表达促进了培养细胞的神经元分化。此外，N-Src异构体在非洲爪哇神经元发育过程中高度调控，N-Src基因敲除抑制神经元分化。目的：了解N-Src异构体在神经系统发育和细胞培养中调节神经元规范和分化的作用。这项基础研究将为了解N-SRCS在神经母细胞瘤和干细胞培养中作为神经分化调节因子的潜力奠定基础。我们为这个问题带来了令人兴奋的专业知识组合。我们已经开发了一种试剂，使我们能够在发育中的胚胎和培养细胞中过度表达并特异性地抑制N-Src的活性。因此，我们处于约克系BIOLOGYWhite Rose BSRC DTP学生申请表SBForm_WR BBSRC DTP，V2.0(2015年9月)第2页进行这项研究的有利位置。及时性：鉴于最近发现选择性剪接在神经发育中发挥关键作用，我们提出的研究Src剪接异构体的神经功能必须被视为时间上的。实验方法1)我们将研究N-Src活性对保守的非洲爪哇神经发生途径中基因表达的影响。在确定了非洲爪哇中关键的N-Src调控元件后，我们将在培养中研究它们在N-Src介导的哺乳动物细胞分化中的作用。2)利用培养中的N-Src分化细胞，我们将获得N-Src调控转录组的整体情况。然后将在发育中的胚胎和细胞中研究已识别的靶标的功能。

(N1-Src and N2-Src) is enriched in neural tissue and high levels of N-Src expression are associated with a favourable prognosis in neuroblastoma, leading to spontaneous differentiation and resolution of the cancer. These observations suggest that N-Srcs regulate neuronal differentiation. Our previous work shows that expression of N1-Src drives neuronal differentiation of cells in culture. Furthermore, N-Src isoform expression is highly regulated during Xenopus neuronal development and N-Src knockdown inhibits neuronal differentiation.Objectives: To understand the role that N-Src isoforms play in regulating neuronal specification and differentiation in the developing nervous system and cell culture. This basic research will underpin efforts to understand the potential of N-Srcs as regulators of neuronal differentiation in neuroblastoma and stem cells in culture.Novelty: The normal function of N-Srcs in neuronal differentiation is poorly understood. We bring an exciting combination of expertise to this problem. We have developed reagents that allow us to overexpress and specifically inhibit N-Src activity in the developing embryo and cells in culture. Thus, we are in a uniquelyUNIVERSITY OF YORKDEPARTMENT OF BIOLOGYWhite Rose BBSRC DTP Studentship Bid FormSBForm_WR BBSRC DTP, V2.0 (Sept 2015) Page 2 of 2favourable position to undertake this study.Timeliness: Given that it has recently become apparent that alternative splicing plays a key role in neural development, our proposed study investigating the neural function of Src splice isoforms must be viewed as timely.Experimental Approaches1) We will investigate the effects of manipulating N-Src activity on expression of genes in the conserved Xenopus neurogenic pathway. Having identified key N-Src regulated elements in Xenopus, we will investigate their function in N-Src mediated differentiation of mammalian cells in culture.2) Using N-Src differentiated cells in culture, we will obtain a global view of the N-Src regulated transcriptome. The function of identified targets will then be investigated in the developing embryo and cell.