Investigation of Mammalian Retinal Neuron Development

摘要 这项建议利用老鼠来研究人类眼部疾病的根本原因。 模特们。我们将重点放在Notch信号通路上，它在多个 哺乳动物组织。特别是，Notch信号调节细胞的增殖、凋亡、 形态变化、分化和干细胞维持。此提案中的实验 将1)阐明Notch信号与Math5之间的上位关系 视网膜神经节细胞(RGC)的神经发生2)探索视网膜多神经元 Rbpj的表型，以及3)定义了Notch配体Deltalike1在 视网膜神经发生。由于Notch信令在开发过程中被广泛使用， 大多数Notch途径基因的小鼠突变已经被创造出来。vbl.使用 靶向缺失小鼠(完全突变和条件等位基因)，我们建议理解 视网膜神经节细胞和视锥细胞对典型Notch信号的要求 杆状感光细胞的形成。一些研究将使用条件性(cre-lox)小鼠 菌株，组织学，免疫组织化学，原位杂交，小鼠胚胎学和 聚合酶链式反应基因分型。其他人将使用人类在体外测试监管关系 视网膜母细胞系和生化分析。这些研究将做出根本性贡献 视网膜前体细胞的生长、形态发生和分化的信息 在所有后生动物的发育过程中。这项工作将使我们更好地理解 视锥-杆状营养不良，视神经发育不全，以及基础 直接与基因或细胞相关的视网膜细胞发育机制 视网膜疗法。这一发现也将对CADASIL的病理学有广泛的帮助 自Notch异常表达以来，Alagille综合征与癌症生物学 途径基因出现在这些疾病中的每一种中。

Abstract This proposal investigates the underlying causes of human ocular diseases using mouse models. We focus on the Notch signaling pathway, which is critically required in multiple mammalian tissues. In particular, Notch signaling regulates proliferation, apoptosis, cell shape changes, differentiation and stem cell maintenance. Experiments in this proposal will 1) elucidate the epistatic relationship between Notch signaling and Math5 during retinal ganglion cell (RGC) neurogenesis 2) explore the multiple retinal neuron phenotypes of Rbpj, and 3) define the requirements of the Notch ligand Deltalike1 during retinal neurogenesis. Because Notch signaling is widely employed during development, mouse mutations in most Notch pathway genes have already been created. Using targeted deletion mice (wholly mutant and conditional alleles), we propose to understand the requirements for canonical Notch signaling during retinal ganglion cell and cone and rod photoreceptor formation. Some studies will employ conditional (cre-lox) mouse strains, histology, immunohistochemistry, in situ hybridization, mouse embryology and PCR genotyping. Others will test regulatory relationships in vitro using a human retinoblast cell line and biochemical assays. These studies will contribute fundamental information retinal progenitor cell growth, morphogenesis and differentiation, which occur throughout all metazoan development. This work will yield a better understanding of cone-rod dystrophies, optic nerve aplasia, hypoplasia, as well as contribute to basic mechanisms of retinal cell development with direct relevance to gene- or cell-based retinal therapies. Findings here will also be widely useful to the pathologies of CADASIL and Alagille Syndromes, and cancer biology, since abnormal expression of Notch pathway genes occurs in each of these diseases.