RUI: Microtubule Regulation in the C. elegans GABAergic neurons

微管在神经元发育和功能中起着复杂的作用，控制突触的生长和稳定以及轴突的生长和运输。细胞已经进化来调节微管的工具库包括微管蛋白折叠辅因子、微管切断蛋白、微管加末端结合蛋白和用于稳定微管阵列的多种类型的蛋白质。微管动力学和与马达蛋白和其他微管相关蛋白的相互作用受到细胞的严格调节，微管和轴突运输的失调是许多神经退行性疾病的病理学的一部分。该项目采用分子遗传学方法在C. elegans研究神经系统发育过程中的微管调节。 该项目的重点是α微管蛋白基因tba-1在轴突和突触发育中的调控。 一个新的等位基因tba-1（ju 89）导致C. elegans GABA能运动神经元，但不破坏早期胚胎细胞分裂或发育。 通过基因筛选，确定了多个抑制突变体（存根基因），以逆转tba-1（ju 89）突变的影响。 因此，TBA-1（JU 89）和抑制子残端突变体提供了一种独特的遗传途径，用于研究正常突触和轴突分化所必需的微管介导的机制，并鉴定导致突触功能障碍和神经元变性的分子机制。目标1：克隆tba-1（ju 89）抑制基因stub-1; AIM 2：研究STUB-1的表达及其与突触前蛋白的共定位; AIM 3：定位stub-2和stub-3; AIM 4：检测stub基因之间的遗传相互作用。该项目将通过为遗传学、细胞生物学和发育神经生物学的学生提供新的研究机会，并为新的跨学科神经科学倡议做出贡献，来加强西方学院的研究和教育。 全面C.西方的线虫研究项目还将提供资源和专业知识，以支持实施新的遗传学和基因组学课程，以及使用线虫RNAi技术和表达转基因GFP标记的蠕虫体内成像的细胞分子生物学入门课程的新实验室模块。

Microtubules play complex roles in neuronal development and function, controlling synapse growth and stabilization as well as axon outgrowth and transport. The arsenal of tools that cells have evolved to regulate microtubules include tubulin folding cofactors, microtubule severing proteins, microtubule plus-end binding proteins and multiple classes of proteins that act to stabilize microtubule arrays. Microtubule dynamics and interactions with motor proteins and other microtubule-associated proteins are tightly regulated by the cell, and dysregulation of microtubules and axon transport is part of the pathology of many neurodegenerative diseases. This project uses a molecular genetic approach in C. elegans to study microtubule regulation during nervous system development. The project focuses on the regulation of an alpha-tubulin gene, tba-1, in axon and synapse development. A novel allele, tba-1(ju89), causes synapse loss in the C. elegans GABAergic motor neurons, but does not disrupt early embryonic cell divisions or development. Multiple suppressor mutants (stub genes) were identified through a genetic screen for mutations that reverse the effects of tba-1(ju89) mutation. tba-1(ju89) and the suppressor stub mutants thus provide a unique genetic avenue to study microtubule-mediated mechanisms that are essential for normal synapse and axon differentiation and to identify molecular mechanisms that lead to synaptic dysfunction and neuron degeneration. AIM 1: clone the tba-1(ju89) suppressor stub-1; AIM 2: study the expression of STUB-1 and its colocalization with presynaptic proteins; AIM 3: map stub-2 and stub-3; and AIM 4: test for genetic interactions between the stub genes. This project will enhance research and education at Occidental College by providing new research opportunities to students in genetics, cell biology and developmental neurobiology and contributing to a new interdisciplinary neuroscience initiative. The presence of a comprehensive C. elegans research program at Occidental will also provide resources and expertise to support the implementation of a new genetics and genomics course and new laboratory modules for the Introductory Cell & Molecular Biology courses using C.elegans RNAi techniques and in vivo imaging of worms expressing transgenic GFP markers.