Identification of factors essential for age-related neuronal health: insights into common mechanisms of neurodegeneration

在对成年斑马鱼进行的基因筛查中，选择了破坏年龄相关神经元的突变 动态平衡，共鉴定出36个基因。这些基因座中的许多以前被显示为关联或 导致人类神经退行性变，如阿尔茨海默氏症、亨廷顿氏症、帕金森氏症和脊髓小脑 共济失调疾病。这项建议描述了解决基因子集的细胞功能的实验 从这个屏幕上识别出具有未知作用机制的。首先，功能突变的完全丧失将 通过在编码序列内进行大的基因组缺失而在这个子集产生。接下来，这些 将通过监测几个细胞应激反应来分析它们对神经元动态平衡的影响 已知在神经退行性变过程中激活的通路，包括内质网应激/未折叠蛋白 反应、氧化应激反应和自噬/有丝分裂吞噬。在对这些研究的预期中，我们小组 已经为这些过程中的每一个建立了基于成像的记者。此外，变种人也将被 在几个敏化遗传菌株的背景下进行评估，在这些菌株中，这些应激反应途径具有 已经妥协了。最后，将通过细胞类型特异性基因来研究突变影响的自主性。 多种神经细胞和非神经细胞类型的破坏。这些研究将提供重要的基线 在包括人类在内的哺乳动物中进行下一步分析的信息和原理 可能是治疗干预的关键目标，以促进神经元健康和预防 神经退行性变。

In a genetic screen conducted in adult zebrafish to select mutations that disrupt age-related neuronal homeostasis, 36 genes were identified. Many of these loci have previously been shown as associated or causative to human neurodegenerations such as Alzheimer's, Huntington's, Parkinson's and Spinocerebellar Ataxia diseases. This proposal describes experiments to address cellular functions of a subset of the genes identified from this screen with unknown mechanisms of action. First, complete loss of function mutations will be generated in this subset though engineered large genomic deletions within coding sequence. Next, these will be analyzed for their impact on neuronal homeostasis by monitoring several cellular stress responsive pathways known to be activated during neurodegeneration including the ER stress/Unfolded Protein Response, Oxidative Stress Response, and Autophagy/Mitophagy. In anticipation of these studies, our group has established imaging-based reporters for each of these processes. In addition, mutants will also be evaluated in the context of several sensitized genetic strains in which these stress-responsive pathways have been compromised. Finally, the autonomy of mutant impact will be investigated through cell-type specific gene disruption in multiple neuronal and non-neuronal cell types. These studies will provide important base-line information and rationale for carrying out next-step analysis in mammalian, including human, homologues that may represent critical targets for therapeutic intervention to promote neuronal health and prevent neurodegeneration.