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Differentiation of dendrites and the analysis of spine- like structures on Lobula Plate Tangential Cells in Drosophila melanogaster

机译:黑腹果蝇Lobula plate Tangential Cells枝状突分化及脊柱样结构分析

摘要

The development of dendrites leads to the establishment of cell-type specific morphology of dendritic trees that eventually determines the way in which synaptic information is processed within the nervous system. The aim of this study was to investigate dendritogenesis of Drosophila motion-sensitive Lobula Plate Tangential Cells (LPTCs) and to understand the role of cytoskeletal molecules in these developmental processes. I employed genetic techniques to obtain fluorescent labeling exclusively in the neurons of interest. In order to visualize the LPTCs confocal imaging was applied.Time point analysis allowed me to follow and describe the phases of LPTC differentiation in the intact Drosophila brain starting from the third instar larva throughout the pupal stages until adulthood. I determined the time when the initial growth of LPTC dendrites starts and showed it to be directional from the beginning. Additionally, I demonstrated that the phase of extensive dendritic growth and branching precedes reorganization processes that lead to establishment of the final architecture of LPTC dendritic trees. In parallel, I attempted to analyze the contribution of actin and tubulin in the shaping of the neurons. In these experiments actin-GFP localized to dendritic termini whereas tubulin-GFP was mainly observed in the primary dendritic branches. These data showed clear similarities between the cytoskeletal organization of LPTCs dendrites and vertebrate neurons. The discovery of the actin enrichment in dendritic termini made me conduct a set of experiments to test if these protrusions are the counterparts of vertebrate spines. I performed a thorough quantitative analysis of spine- like protrusions present on LPTC dendrites. Morphological features like the density and shape of the LPTC spine- like protrusions appeared to be comparable to hippocampal spines. Using immunohistochemical methods I demonstrated that LPTC spine-like protrusions are sites of synaptic contacts. The ultrastructural analysis supported the immunohistochemical data and showed that synaptic transmission takes place at the LPTC spine-like protrusions. Next, I tried to genetically modify these structures by generating LPTC mutant for genes which have vertebrate homologues known to alter spine morphology. I showed that dRac1 can modulate significantly the LPTC spine-like structure density. Finally, I tried to check if Drosophila LPTC spine-like structures are motile. To conclude, I showed an initial description of LPTC dendritogenesis and the subcellular localization of actin and tubulin in these neurons. The actin enriched spine-like structures detected on the LPTC dendrites are sites of synaptic contacts, thus resemble vertebrate spines.
机译:树突的发展导致树突树的细胞类型特异性形态的建立,最终决定了神经系统中突触信息的处理方式。这项研究的目的是调查果蝇运动敏感性小叶板切向细胞(LPTC)的树突形成,并了解细胞骨架分子在这些发育过程中的作用。我采用遗传技术仅在感兴趣的神经元中获得荧光标记。为了使LPTCs可视化,应用了共聚焦成像。时间点分析使我能够追踪并描述完整的果蝇大脑中LPTC分化的阶段,从整个throughout期的第三龄幼虫开始直至成年。我确定了LPTC树突的初始生长开始的时间,并表明它从一开始就是定向的。此外,我证明了树突状生长和分支扩展的阶段先于重组过程,最终导致建立LPTC树突状树的最终架构。同时,我尝试分析肌动蛋白和微管蛋白在神经元塑造中的作用。在这些实验中,肌动蛋白-GFP定位于树突状末端,而微管蛋白-GFP主要在初级树突状分支中观察到。这些数据显示LPTC树突和脊椎动物神经元的细胞骨架组织之间明显相似。在树突状末端中肌动蛋白富集的发现使我进行了一组实验,以测试这些突起是否与脊椎动物的棘突相对应。我对LPTC树突上的脊柱状突起进行了彻底的定量分析。 LPTC脊柱状突起的密度和形状等形态特征似乎与海马棘相当。使用免疫组织化学方法,我证明了LPTC脊柱状突起是突触接触的部位。超微结构分析支持了免疫组织化学数据,并表明突触传递发生在LPTC脊柱样突起处。接下来,我试图通过产生LPTC突变体来遗传修饰这些结构,这些突变体的基因具有已知会改变脊柱形态的脊椎动物同源基因。我表明dRac1可以显着调节LPTC脊柱状结构密度。最后,我尝试检查果蝇LPTC脊柱状结构是否活动。总而言之,我对LPTC树突生成以及这些神经元中肌动蛋白和微管蛋白的亚细胞定位进行了初步描述。 LPTC树突上检测到的富含肌动蛋白的脊柱样结构是突触接触的部位,因此类似于脊椎动物的棘突。

著录项

  • 作者

    Koper Ewa;

  • 作者单位
  • 年度 2007
  • 总页数
  • 原文格式 PDF
  • 正文语种 {"code":"de","name":"German","id":7}
  • 中图分类
  • 入库时间 2022-08-20 21:04:21

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