SPINE MORPHOLOGY IN AN ANIMAL MODEL OF FRAGILE X SYNDROME

脆性 X 综合征动物模型的脊柱形态

• 批准号: 7358036
• 负责人: WILLIAM T GREENOUGH
• 金额: $ 3.46万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-05-01 至 2007-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7358036
• 关键词: SPINE; MORPHOLOGY; ANIMAL; MODEL; FRAGILE

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Golgi-impregnation studies of human autopsy tissue and of a mouse model of fragile X mental retardation syndrome suggest that dendritic spines have an underdeveloped appearance ¿ a tendency towards elongated, tortuous spine morphology. Fragile X mental retardation, a genetically-linked disorder, results from the inability to produce the protein FMRP. The lack of FMRP expression has been associated with altered dendritic spine morphology in human post-mortem tissue and a mouse model of the disorder. Moreover, stability of spine shape has been shown to be dependent upon the filamentous form of actin, F-actin. In this study we compared spine morphology in mice with knockout (KO) of the FMRP gene to that of wild-type (WT) controls. We used of phalloidin conjugated flurophores followed by photooxidation for correlative analysis of F-actin stained spines at the light and electron microscope levels and advanced structural imaging tomography after cell filling with Lucifer yellow and photoconversion. All tomography was performed on the 3MeV UHVEM microscope in Osaka Univerisity on sections 4-5 ¿m thick. The thicker sections were necessary in order to sample a reasonable length of dendrite and to capture larger spines in their entirety. We report three primary observations. First, pedunculated spines in KO mice are much more likely to stain for F-actin than in WT. Second, the mean spine length of F-actin stained spines in KO mice is longer than WT. Third, tomography data enable us to better correlate these findings, and others, to more accurately characterize the morphological phenotype expressed in this disorder. These findings are consistent with previous reports using Golgi impregnation while those from the tomography offer tremendous potential to accurately and efficiently elucidate the morphological profile of this dis

该子项目是利用NIH/NCRR资助的中心赠款提供的资源的许多研究子项目之一。子项目和研究者（PI）可能从另一个NIH来源获得主要资金，因此可以在其他CRISP条目中表示。所列机构为中心，不一定是研究者所在机构。对人类尸检组织和脆性X染色体智力低下综合征小鼠模型的高尔基体浸渍研究表明，树突棘具有不发达的外观-倾向于细长，曲折的棘形态。脆性X智力低下是一种遗传性疾病，是由于不能产生蛋白质FMRP而引起的。FMRP表达的缺乏与人类死后组织和该疾病的小鼠模型中树突棘形态的改变有关。此外，棘形状的稳定性已被证明是依赖于丝状形式的肌动蛋白，F-肌动蛋白。在这项研究中，我们比较了脊柱形态与敲除（KO）的FMRP基因的小鼠野生型（WT）的控制。我们使用鬼笔环肽共轭荧光团，然后光氧化的F-肌动蛋白染色的棘在光镜和电子显微镜水平和先进的结构成像断层扫描后，细胞填充与荧光黄和光转换的相关性分析。所有断层扫描均在大坂大学的3 MeV UHVEM显微镜上进行，切片厚度为4-5 μ m。较厚的切片是必要的，以便对合理长度的树突进行采样，并完整地捕获较大的棘。我们报告三个主要观察。首先，KO小鼠的有蒂棘比WT小鼠更容易对F-肌动蛋白染色。第二，KO小鼠中F-肌动蛋白染色的棘的平均棘长度比WT长。第三，断层扫描数据使我们能够更好地将这些发现与其他发现相关联，以更准确地表征这种疾病中表达的形态表型。这些发现与以前使用高尔基体浸渍的报道一致，而断层扫描的结果为准确有效地阐明这种疾病的形态学特征提供了巨大的潜力。