Neuronal Mechanisms of Peroxisomal Biogenesis Defects in Drosophila

描述(由申请人提供)：该提案描述了一项为期五年的有指导的实验室培训经验，旨在引导您在与临床相关的基础科学领域取得独立的学术生涯。申请人拥有医学博士学位，并已完成儿科学专业培训和委员会认证，目前正在完成医学遗传学的子专业培训。职业发展计划包括一段时间的有指导的研究培训，其中将包括学习研究技术和概念，并辅之以教学培训、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，并使申请者有机会在学习电子显微镜和电生理学等强大技术方面得到指导。本研究旨在通过对果蝇过氧化物体生物发生的研究，在分子水平上提高我们对过氧化物体生物发生障碍的认识。过氧化体是真核生物中普遍存在的细胞器，由Pex基因编码的一组进化保守的蛋白质产生。人类Pex基因座的突变会导致Peroxisomal生物发生障碍(PBD)，这是一种具有毁灭性神经后果的疾病。PBD的神经系统并发症已有特征，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体生物发生缺陷的了解。人们对果蝇的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，只有很少的工具可用于研究果蝇的过氧化物体，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、标记的基因组结构以及表征过氧化物体结构和功能的分析。我们将通过检验过氧化物体丢失影响线粒体功能的假设来探索过氧化物体与其他细胞器的相互作用。最后，我们将以我们的初步数据为基础，通过定义缺陷的过氧化体生物发生导致突触传递中的胞外缺陷来显示pex16 P元件插入突变体的电生理缺陷。这项研究将使人们对过氧化物体生物发生对神经系统的影响有更广泛的了解。这可能对患有过氧素体疾病的患者有临床意义。这项研究也将在一个专门培训申请者作为独立科学家进一步从事这项研究的环境中进行。 公共卫生相关性：这项研究研究了一组疾病，在这些疾病中，患者缺乏一种名为过氧化物体的重要细胞器，导致身体化学问题。研究人员使用具有类似基因改变的果蝇来研究由于缺乏过氧化物酶体而导致的问题。这项研究可能会让人们更好地了解这些没有有效治疗方法的疾病。

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. PUBLIC HEALTH RELEVANCE: This research studies a group of diseases in which the patient lacks an important cellular organelle named peroxisomes leading to problems in body chemistry. Researchers use fruit flies with similar genetic alterations to study the problems that result from lacking peroxisomes. The research could lead to a better understanding of these diseases for which there are no effective treatments.