Effects of inflammation on developing glia

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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. We have developed a model of intrauterine inflammation using pregnant rats that (i) causes damage to developing oligodendrocytes and (ii) demonstrates a cytokine response in the developing brain. We hypothesize that cytokines, released in response to the inflammatory stimulus, are responsible for damage to developing oligodendrocytes during intrauterine inflammation. We propose a 5-year, multidisciplinary research and training program that focuses on testing this hypothesis in two related experimental models of inflammation, including transgenic mice. To this end, we will determine the effects of experimental intrauterine inflammation on the proliferation and differentiation of cells of the oligodendrocyte lineage in rats. We will characterize the cytokine response in fetal brain and placenta at the protein and mRNA level. We will develop a similar inflammatory model in mice and use transgenic mice lacking TNF receptors to determine the role of TNF-alpha in the damage to developing oligodendrocytes. We 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, we will compare expression of genes involved in the inflammatory cascade after our experimental stimulus. This research grant will extend the investigator'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 the investigator with sophisticated 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.