Characterization of primary cilia during the differentiation of retinal ganglion cells in the zebrafish.
Characterization of primary cilia during the differentiation of retinal ganglion cells in the zebrafish.
复制标题
DOI：
10.1186/s13064-016-0064-z
复制
发表时间：
2016-04-06
影响因子：
3.6
通讯作者：
Zolessi FR
中科院分区：
文献类型：
--
作者：
Lepanto P;Davison C;Casanova G;Badano JL;Zolessi FR
视网膜神经节细胞（Retinal ganglion cell，RGC）在体内的分化是一个高度定型的过程，可能是细胞类型特异性转录因子和组织来源的信号传导因子相互作用的结果。初级纤毛作为细胞中的信号传导枢纽，可能在此过程中发挥作用，但其在RGC生成过程中的存在和定位及其对细胞分化过程的贡献先前尚未在体内评估。在这项工作中，我们分析了初级纤毛在体内的分布，使用激光扫描共聚焦显微镜，以及其主要的超微结构特征，在视网膜组织发生的早期阶段，在斑马鱼，周围的时间RGC生成和初始分化。此外，我们敲除ift 88和elipsa，这两个基因在纤毛的生成和维持中起着重要作用，这种处理导致细胞器尺寸的普遍减小。通过转基因或免疫标记胚胎的共聚焦显微镜评估对视网膜发育和RGC分化的影响。我们的研究结果表明，视网膜神经上皮细胞有一个apicallylocalized初级纤毛通常从顶膜突出。我们还发现了一个小比例的亚顶端纤毛，之前和期间的神经。这个细胞器也存在于顶端的位置在成神经细胞在顶端的过程中收缩和dendritogenesis，虽然这些阶段之间的纤毛出现高度动态的存在和位置。破坏纤毛导致视网膜祖细胞增殖减少和神经视网膜体积减少。此外，虽然RGC层的形成相对于无长突和感光细胞层优先减少，但视网膜组织发生总体延迟。这些结果表明，初级纤毛在早期视网膜分化过程中表现出高度动态的行为，并且它们是视网膜祖细胞的增殖和存活以及神经元生成（特别是RGC）所需的。本文的在线版本（doi：10.1186/s13064-016-0064-z）包含补充材料，可供授权用户使用。
Retinal ganglion cell (RGC) differentiation in vivo is a highly stereotyped process, likely resulting from the interaction of cell type-specific transcription factors and tissue-derived signaling factors. The primary cilium, as a signaling hub in the cell, may have a role during this process but its presence and localization during RGC generation, and its contribution to the process of cell differentiation, have not been previously assessed in vivo. In this work we analyzed the distribution of primary cilia in vivo using laser scanning confocal microscopy, as well as their main ultrastructural features by transmission electron microscopy, in the early stages of retinal histogenesis in the zebrafish, around the time of RGC generation and initial differentiation. In addition, we knocked-down ift88 and elipsa, two genes with an essential role in cilia generation and maintenance, a treatment that caused a general reduction in organelle size. The effect on retinal development and RGC differentiation was assessed by confocal microscopy of transgenic or immunolabeled embryos. Our results show that retinal neuroepithelial cells have an apically-localized primary cilium usually protruding from the apical membrane. We also found a small proportion of sub-apical cilia, before and during the neurogenic period. This organelle was also present in an apical position in neuroblasts during apical process retraction and dendritogenesis, although between these stages cilia appeared highly dynamic regarding both presence and position. Disruption of cilia caused a decrease in the proliferation of retinal progenitors and a reduction of neural retina volume. In addition, retinal histogenesis was globally delayed albeit RGC layer formation was preferentially reduced with respect to the amacrine and photoreceptor cell layers. These results indicate that primary cilia exhibit a highly dynamic behavior during early retinal differentiation, and that they are required for the proliferation and survival of retinal progenitors, as well as for neuronal generation, particularly of RGCs. The online version of this article (doi:10.1186/s13064-016-0064-z) contains supplementary material, which is available to authorized users.