Effects of inflammation on developing glia

描述（由申请人提供）：脑瘫（CP）是影响成千上万儿童的最常见的发育疾病之一。脑瘫是由大脑白质正常发育受到干扰引起的。孕妇的宫内炎症可能导致高达12%的CP病例。动物模型的建立对于确定炎症与CP之间关联的机制至关重要。该候选人利用怀孕大鼠建立了宫内炎症模型，该模型要么对发育中的少突胶质细胞造成损伤，要么在发育中的大脑中表现出细胞因子反应。据推测，在宫内炎症期间，响应炎症刺激而释放的细胞因子是导致发育中的少突胶质细胞受损的原因。候选人提出了一个为期五年的多学科研究和培训计划，重点是在两种相关的炎症实验模型中测试这一假设，包括转基因动物。为此，他将确定实验性宫内炎症对大鼠少突胶质细胞系细胞增殖和分化的影响。该候选人将在蛋白质和mRNA水平上表征胎儿脑和胎盘的细胞因子反应。他将在小鼠中建立类似的炎症模型，并使用缺乏TNF受体的转基因动物来确定TNF-(在发育中的少突胶质细胞损伤中的作用。他将使用一种转基因小鼠品系，该品系在少突胶质细胞特异性启动子下游插入增强型绿色荧光蛋白的报告基因，在炎症刺激后通过荧光激活细胞分选（FACS）从这些细胞中纯化少突胶质前体细胞。通过提取RNA，他将比较实验刺激后炎症级联反应相关基因的表达。该职业奖将通过在特定技术（RT-PCR, Western blot，使用基因微阵列的基因表达谱和报告标记细胞的FACS分选）方面的指导和培训来扩展候选人的能力。此外，这将为他提供分析和解释数据所需的技能，以解决从遗传到细胞水平的炎症复杂病理生理过程。神经科学方面的指导以及智力迟钝和发育障碍研究中心的资源将支持这些明确的培训目标。

DESCRIPTION (provided by applicant): Cerebral palsy (CP) is one of the most common developmental diseases affecting thousands of children. CP is caused by disturbances of the normal development of the cerebral white matter. Intrauterine inflammation in pregnant women may be responsible for up to 12% of the cases of CP. Development of animal models is essential to determine the mechanisms responsible for the association between inflammation and CP. The candidate has developed a model of intrauterine inflammation using pregnant rats that either causes damage to developing oligodendrocytes or demonstrates a cytokine response in the developing brain. It is hypothesized that cytokines, released in response to the inflammatory stimulus, are responsible for damage to developing oligodendrocytes during intrauterine inflammation. The candidate proposes a five-year, multidisciplinary research and training program that focuses on testing this hypothesis in two related experimental models of inflammation, including transgenic animals. To this end, he will determine the effects of experimental intrauterine inflammation on the proliferation and differentiation of cells of the oligodendrocyte lineage in rats. The candidate will characterize the cytokine response in fetal brain and placenta at the protein and mRNA level. He will develop a similar inflammatory model in mice and use transgenic animals lacking TNF receptors to determine the role of TNF-( in the damage to developing oligodendrocytes. He will use a transgenic mouse strain that has a reporter gene for enhanced green fluorescent protein inserted downstream of an oligodendrocyte-specific promoter to purify oligodendrocyte precursor cells from these cells by fluorescence-activated cell sorting (FACS) after the inflammatory stimulus. By extracting RNA, he will compare expression of genes involved in the inflammatory cascade after the experimental stimulus. This career award will extend the candidate's capabilities through mentorship and training in specific techniques (RT-PCR, Western blot, gene expression profiling using gene microarrays and FACS sorting of reporter-labeled cells). In addition, it will provide him with skills in the analysis and interpretation of data required to address the complex pathophysiological processes of inflammation from the genetic to cellular level. Mentorship in neuroscience as well as the resources of the Mental Retardation and Developmental Disabilities Research Center will support these well-defined training objectives.