GROWTH FACTORS IN MAMMALIAN EYE DEVELOPMENT

哺乳动物眼睛发育的生长因素

• 批准号: 2670116
• 负责人: Jeffrey L Bennett
• 金额: $ 15.26万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-07-01 至 2003-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2670116
• 关键词: biological signal transduction; cell differentiation; confocal scanning microscopy; cornea; corneal epithelium; developmental genetics; eye; fluorescence microscopy; gene targeting; growth /development; growth factor receptors; immunocytochemistry; laboratory mouse; lens; mammalian embryology; neurotrophic factors; retina; uvea ciliary body

DESCRIPTION: (Candidate's Abstract) The ultimate aim of this proposal is to formulate a multifaceted program which will guide my development as an independent scientist. This goal is to be accomplished through a mixture of basic research and educational activities. My research will be focused on determining the biologic function of the neurotrophins brain-derived neurotrophic factor (BDNF) and neurotrophin-3 (NT-3) in the development and maintenance of the mammalian eye. These endeavors will be complemented by extramural workshops, instruction in ocular pathology, and participation at scientific meetings. I have spent the past several years completing my doctoral research and professional training. I am now prepared to embark on a long-term career in academic neuro-ophthalmology, combining clinical practice with basic research. The University of Colorado will provide me with an ideal environment in which to accomplish my immediate and long-term goals. The combined faculties and resources of the Department of Neurology, Ophthalmology, and Molecular, Cellular, and Developmental Biology are particularly well suited for providing me with training in nervous system and eye development, mouse genetics, histochemistry, and transgenic and embryonic stem cell technology. Under the research mentorship of Dr. Kevin Jones, I will investigate the biologic roles of BDNF and NT-3 in the neuroretina and anterior segment of the eye. A multidisciplinary approach will address questions in three main areas. First, the cell specific expression of BDNF and NT-3 and their respective receptors. TrkB and TrkC, will be examined in the neuroretina, lens, ciliary body, and cornea. We will identify the specific cells expressing BDNF, NT-3, TrkB, and TrkC using dual-label immunohistochemistry and confocal microscopy. These results will suggest possible functions for these neurotrophins in the eye. Second, the biologic functions of BDNF and NT-3 in the neuroretina will be investigated using subregion-restricted gene knockout technology. Mice will be generated which lack BDNF and NT-3 in the neuroretina. To achieve this, we will use Cre/loxP-mediated recombination restricted by either the Six3 or Brn-3b promoter. Cre recombinase will be expressed either as a transgene or as a dicistronic message using an intervening ribosome entry sequence (IRES). Third, the biologic role of BDNF and NT-3 in the anterior segment of the eye will be characterized following our recent finding of regulated neurotrophin expression in the ciliary body epithelium, lens epithelium, and cornea. Restricted gene knockouts will be constructed using dicistronic Cre expression driven from the endogenous aquaporin-1 gene. These studies should provide novel insight into the molecular genetics of eye development, potentially broaden our understanding of degenerative eye disorders and open new avenues for their treatment with growth factors.

描述：（候选人摘要）本提案的最终目的 是制定一个多方面的计划来指导我的发展 作为一个独立的科学家。这一目标将通过一个 基础研究和教育活动的结合。我的研究将 集中于确定神经营养因子的生物学功能 脑源性神经营养因子（BDNF）和神经营养素-3（NT-3）在 哺乳动物眼睛的发育和维持。这些努力 将辅以校外讲习班，在视觉教学 病理学和参加科学会议。 我花了几年时间完成了我的博士研究， 专业训练我现在准备开始一个长期的职业生涯 在学术神经眼科，结合临床实践与基础 research.科罗拉多大学将为我提供一个理想的 在这样的环境中，我可以实现我的短期和长期目标。的 结合神经病学系的师资和资源， 眼科学，分子，细胞和发育生物学 特别适合为我提供神经训练， 系统和眼睛发育，小鼠遗传学，组织化学， 转基因和胚胎干细胞技术。 在凯文·琼斯博士的研究指导下，我将调查 BDNF和NT-3在神经视网膜和眼前节中的生物学作用 的眼睛。多学科方法将解决三个方面的问题 主要领域。 第一，BDNF和NT-3的细胞特异性表达及其与细胞凋亡的关系。 各自的受体。TrkB和TrkC将在 神经视网膜、透镜、睫状体和角膜。我们将确定 使用双标记表达BDNF、NT-3、TrkB和TrkC的特异性细胞 免疫组织化学和共聚焦显微镜。这些结果表明 这些神经营养素在眼睛中的可能功能。 第二，BDNF和NT-3在神经视网膜中的生物学功能将进一步被研究。 使用亚区限制性基因敲除技术进行研究。 将产生在神经视网膜中缺乏BDNF和NT-3的小鼠。到 为了实现这一点，我们将使用Cre/loxP介导的重组， Six3或Brn-3b启动子。Cre重组酶将被表达 作为转基因或作为使用插入的双顺反子信息， 核糖体进入序列（IRES）。 第三，BDNF和NT-3在眼前节中的生物学作用， 眼睛将根据我们最近的发现， 睫状体上皮，透镜上皮， 和角膜。限制性基因敲除将使用 由内源性水通道蛋白-1基因驱动的双顺反子Cre表达。 这些研究将为分子遗传学提供新的见解 眼睛的发展，潜在地扩大了我们对 退行性眼部疾病，并为他们的治疗开辟了新的途径， 生长因子