Growth & Gene Expression in Primary Sensory Neurons

我的长期目标是成为一名独立的研究人员，为我们对中枢神经系统（CNS）创伤的理解做出重大贡献，特别是脊髓创伤。我的博士前工作重点是利用生化、分子和行为结果研究挫伤脊髓损伤后的初始兴奋毒性事件和慢性中枢性疼痛的发展。为了成为一名独立的脊髓损伤研究者，我需要对中枢神经系统损伤后的神经再生有更深入的了解。在这项研究中，我将通过评估损伤后生长相关基因的表达来研究为什么中枢神经系统不能启动新的轴突生长。这需要学习新的技术，如原位杂交、病毒载体传递、轴突生长的体外和体内检测。神经再生模型和一种新的脊髓损伤模型（背柱病变）在这个奖学金期间接受的训练将使我获得独立所需的知识和技术技能Richard J . K.itz麻醉研究教授研究兴趣/领域神经再生/疼痛的神经生物学描述（请不要超过所提供的空间）中枢神经系统（CNS）损伤后的功能丧失是由于中枢神经系统内轴突无法再生造成的，这与周围神经系统（PNS）形成对比，周围神经系统会发生再生尽管多年的研究，负责PNS能力的机制，但失败了。最近，基因表达技术的进步使得使用高密度DNA微阵列同时筛选和鉴定数千种基因表达谱成为可能。使用这项新技术，似乎调节神经元生长的基因已经被鉴定出来。因此，需要对这些推测的生长相关基因（GAGs）进行进一步的表征。本研究的目的是利用背根神经节（DRG）神经元的独特特性来验证通过微阵列分析鉴定的推测的GAGs的所有特征。(3)外周损伤后表达上调，(4)中枢病变后不表达或下调。GAGs将通过slot、Northern和Western blots、免疫细胞化学、满足生长促进GAG标准的基因将在体外和体内进行检查，以确定其在中枢神经系统损伤后调节轴突生长和再生的能力。这些研究结果将增加我们对中枢再生失败原因的理解，并可能为治疗中枢神经系统损伤提供新的治疗机会======================================== 节结束 ===========================================

18 GOALS FOR FELLOWSHIP TRAINING AND CAREER My long-term goal is to become an independent researcher and make significant contributions to our understanding of central nervous system (CNS) trauma, specifically trauma to the spinal cord My predoctoral work focused on the initial excitotoxic events and development of chronic central pain following contusion spinal cord injury using biochemical, molecular, and behavioral outcomes However, in order to become an independent investigator of spinal cord injury, I need a greater knowledge of nerve regeneration after CNS injury During this fellowship I will examine why the CNS fails to initiate new axonai growth by evaluating expression of growth associated genes after injury This requires learning new techniques, such as in situ hybridiazation, viral vector delivery, in vitro and in rive assays of axonal growth, models of nerve regeneration and a new model of spinal cord injury (dorsal column lesion) The training received during this fellowship will allow me to acquire the knowledge and technical skills I need to become independent SPONSOR 19 NAMEANDDEGREE(S) Clifford J Woolf, MD, Phi) 20 POSITION/RANK Richard J K.itz Professor of Anesthesia Research 21 RESEARCHINTERESTS/AREAS Neural Regeneration / Neurobiology of Pain DESCRIPTION (Do not exceed space provided) Loss of function following injury to the central nervous system (CNS) is due to a failure of axons within the CNS to re-grow This is in contrast to the peripheral nervous system (PNS) where re-growth occurs Despite years of study, the mechanisms resgonsible for the ability of the PNS, but failure oft.he CNS to regenerate remain poorly understood Recently, advances in gene expression technology have made it possible to screen and identify thousands of gene expression profiles simultaneously using high density DNA microarrays Using this new technology, genes that appear to regulate neuronal growth have been identified, however, identification by microarrays is inadequate evidence to draw conclusions about functional significance Further characterization of these putative growth associated genes (GAGs) is therefore reqnired The aim of this study is to use the unique properties of dorsal root ganglion (DRG) neurons to validate mad characterize putative GAGs identified by microarry analysis Expression profiles of putative DRG GAGs that have the following characteristics will be examined (i) upregulated during development, (it) down regulated or not expressed in adult, (iii) upregulated after peripheral injury, and (iv) not expressed or down regulated after a central lesion The expression profiles of putative DR(; GAGs will be validated by slot, Northern, and Western blots, immunocytochemistry, and by in situ hybridization to determine cellular localization Genes that fulfill the criteria for a growth promoting GAG will be examined in vitro and in rive for their ability to regulate axonal growth and regeneration after CNS injury These restdts will increase our understanding of the reasons for failure of central regeneration and may offer novel therapeutic opportunities to treat CNS injuries PHS 416-1 (Rev 12/98) Form Page 2 BB CC Individual NRSA Application Table of Contents ========================================Section End===========================================