Growth & Gene Expression in Primary Sensory Neurons

生长

• 批准号: 6584943
• 负责人: CHARLES D MILLS
• 金额: $ 4.16万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-01-01 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6584943
• 关键词: developmental genetics; developmental neurobiology; gene expression; genetic regulation; immunocytochemistry; in situ hybridization; laboratory rat; microarray technology; nerve injury; nervous system regeneration; neurogenetics; neurons; neurotrophic factors; northern blottings; oligonucleotides; postdoctoral investigator; spinal ganglion; western blottings

DESCRIPTION (provided by applicant): 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 responsible for the ability of the PNS, but failure of the 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 required The aim of this study is to use the unique properties of dorsal root ganglion (DRG) neurons to validate and characterize putative GAGs identified by microarray analysis. Expression profiles of putative DRG GAGs that have the following characteristics will be examined (i) upregulated during development, (ii) 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 DRG 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 vivo for their ability to regulate axonal growth and regeneration after CNS injury. These results will increase our understanding of the reasons for failure of central regeneration and may offer novel therapeutic opportunities to treat CNS injuries.

描述（由申请人提供）：中枢神经系统（CNS）损伤后的功能丧失是由于CNS内轴突无法再生。这与周围神经系统（PNS）形成对比，周围神经系统（PNS）发生再生长。尽管多年的研究，负责PNS的能力，但CNS再生失败的机制仍然知之甚少。近年来，基因表达技术的发展使得使用高密度DNA微阵列同时筛选和鉴定数千个基因表达谱成为可能。使用这项新技术，人们已经识别出了似乎调节神经元生长的基因，然而，通过微阵列进行识别并不足以得出有关功能意义的结论。因此，需要进一步表征这些推定的生长相关基因（GAG）。本研究的目的是使用背根神经节（DRG）神经元的独特性质来验证和表征通过微阵列分析鉴定的推定的GAG。将检查具有以下特征的推定DRG GAG的表达谱：（i）在发育期间上调，（ii）在成体中下调或不表达，（iii）在外周损伤后上调，和（iv）在中枢损伤后不表达或下调。将通过狭缝、北方和Western印迹、免疫细胞化学和原位杂交来验证推定DRG GAG的表达谱，以确定细胞定位。将在体外和体内检查满足生长促进GAG标准的基因调节CNS损伤后轴突生长和再生的能力。这些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。