Neuronal Mechanisms of Peroxisomal Biogenesis Defects in Drosophila

描述（由申请人提供）：该提案描述了为期五年的指导实验室培训经历，旨在引导临床相关基础科学的独立学术生涯。申请人拥有医学博士学位，并已完成儿科专业培训和委员会认证，目前正在完成医学遗传学的分专业培训。职业发展计划包括一段时间的指导研究训练，其中包括学习研究技术和概念，辅以教学训练、研讨会、实验室会议、期刊俱乐部、国家和国际会议、咨询委员会和与导师的会议。研究环境提供了最好的智力环境和最好的技术，并为申请人提供了学习电子显微镜和电生理学等强大技术的指导机会。本研究旨在通过关注果蝇过氧化物酶体的生物发生，在分子水平上提高我们对过氧化物酶体生物发生障碍的理解。过氧化物酶体是真核生物中普遍存在的细胞器，由一组由pex基因编码的进化上保守的蛋白质产生。人类过氧化物酶体基因座突变导致过氧化物酶体生物发生障碍（PBD），这是一种具有破坏性神经系统后果的疾病。PBD的神经系统并发症已被描述，但其机制尚不清楚。果蝇提供了一个很好的模型系统来增加我们对过氧化物酶体生物发生缺陷的认识。我们对果蝇的pex基因知之甚少。我们选择了pex2和pex16进行分析。由于这种方法的新颖性，研究果蝇过氧化物酶体的工具非常少，因此我们将生成额外的工具，以允许对pex2和pex16进行精确和彻底的分析，包括零等位基因，标记基因组结构，以及表征过氧化物酶体结构和功能的分析。我们将通过测试过氧化物酶体损失影响线粒体功能的假设来探索过氧化物酶体与其他细胞器的相互作用。最后，我们将在初步数据的基础上，通过定义过氧化物酶体生物发生缺陷导致突触传递中胞外缺陷的机制，揭示pex16 p元素插入突变体的电生理缺陷。这项研究将对过氧化物酶体生物发生对神经系统的影响有更广泛的了解。这可能对过氧化物酶体疾病患者具有临床意义。这项研究也将在一个致力于培养申请人作为独立科学家进一步从事这项研究的环境中进行。

DESCRIPTION (provided by applicant): The proposal describes a five-year mentored laboratory training experience designed to lead to an independent academic career in clinically-relevant basic science. The applicant holds an M.D. degree, and has completed specialty training and board certification in pediatrics and is currently completing sub-specialty training in Medical Genetics. The career development plan includes a period of mentored research training which will include learning research techniques and concepts supplemented by didactic training, seminars, lab meetings, journal clubs, national and international meetings, an advisory committee and meetings with the mentor. The research environment provides the best intellectual environment and the best technology available and gives the applicant the opportunity to be guided in learning powerful techniques such as electron microscopy and electrophysiology. The research seeks to improve our understanding of peroxisomal biogenesis disorders at the molecular level by focusing on peroxisomal biogenesis in Drosophila. Peroxisomes are ubiquitous organelles in eukaryotes, generated by a set of evolutionarily conserved proteins encoded by the pex genes. Mutations in pex loci in humans lead to Peroxisomal Biogenesis Disorders (PBD), diseases with devastating neurologic consequences. The nervous system complications of PBD have been characterized but their mechanism is not known. Drosophila provides a good model system to add to our knowledge of peroxisomal biogenesis defects. Very little is known about the pex genes in Drosophila. We have selected pex2 and pex16 for analysis. Because of the novelty of this approach very minimal tools are available to study peroxisomes in Drosophila, we will therefore generate additional tools to allow for a precise and thorough analysis of pex2 and pex16 including null alleles, tagged genomic constructs, and assays for the characterization of peroxisome structure and function. We will explore the interaction of peroxisomes with other organelles by testing the hypothesis that peroxisomal loss affects mitochondrial function. Finally, we will build on our preliminary data showing electrophysiologic defects in pex16 P-element insertion mutants by defining the mechanisms by which defective peroxisomal biogenesis lead to exocytic defects in synaptic transmission. This research will create a broader understanding of the effect of peroxisomal biogenesis on the nervous system. This could have clinical implications for patients with peroxisomal disorders. This research will also occur in an environment dedicated to training the applicant to pursue this research further as an independent scientist.