Tau Isoform Expression in FTDP-17

FTDP-17 中的 Tau 同工型表达

• 批准号: 6896373
• 负责人: VIVIANNA M VAN DEERLIN
• 金额: $ 13.12万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-06-15 至 2006-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6896373
• 关键词: RNA splicing; cell population study; chromosomes; complementary DNA; dissection; gene expression; gene mutation; glia; human tissue; messenger RNA; microarray technology; neurogenetics; neurons; phenotype; phosphorylation; polymerase chain reaction; protein isoforms; protein structure function; tau proteins; tissue /cell culture

DESCRIPTION (provided by applicant): Filamentous aggregates of hyperphosphorylated tau are the signature brain lesions of frontotemporal dementia with parkinsonism linked to chromosome 17 (FTDP 17), an inherited tauopathy with diverse, phenotypes caused by different tau gene mutations. Tau is a microtubule (MT) binding protein that promotes tubulin polymerization into MTs and stabilizes MTs. The adult human brain contains six tau isoforms, half with 3 3R isoforms) and half with 4 Cterminal MTbinding repeats (4R isoforms) generated by alternatively splicing of exon 10 (E 10). Tau gene mutations cause FTDP 17 by impairing E 10 alternative splicing or tau functions. A puzzling aspect of the FTDP 17 syndromes is that different tau mutations damage selected subtypes of neurons and glia. Our first hypothesis is that this selective vulnerability may reflect cell type specific perturbations of tau isoforms to cause varied phenotypes. To test this hypothesis, we will determine mRNA expression profiles of tau isoforms in normal brain cell subpopulations. Although the ratio of 3R to 4R tau is 1:1 in normal human brain, it has never been determined in subpopulations of neurons and glia. In, Aim 1, we will microdissect neurons and glia from paraffinembedded tissue sections of control brains, perform linear amplification on extracted RNA followed by quantitative realtime RTPCR to measure the relative levels of each tau isoform mRNA. In Aim 2, we will similarly study the same neuronal and glial cell populations in FTDP 17 brains. Correlation of these data with disease phenotypes will clarify mechanisms of FTDP1 7. Since FTDP1 7 mutations produce different phenotypes in the same kindred, a second hypothesis proposes that altered expression of a second gene, which interacts with the tau gene or protein, influences development of phenotypic manifestations of FTDP 17 in different affected family members. Aim 3 tests this hypothesis by examining the differential expression of candidate genes (i.e. those involved in splicing, RNA stability, tau function, other cellular processes) between affected and unaffected FTDP 17 brain regions and control brains using custommade cDNA macroarrays. The completion of these Aims will advance understanding of FTDP 17 and related tauopathies, and may provide new targets for diagnosis and therapeutics.

性状（由申请人提供）： 过度磷酸化的tau蛋白是额颞叶的标志性脑损伤 与17号染色体（FTDP 17）相关的帕金森痴呆症，一种遗传性帕金森病， 具有由不同tau基因突变引起的多种表型的tau蛋白病。Tau 是微管（MT）结合蛋白，其促进微管蛋白聚合成 MT和稳定MT。成年人的大脑含有六种tau亚型， 有3个3R亚型）和一半有4个C末端MT结合重复序列（4R亚型） 由外显子10（E10）的选择性剪接产生。Tau基因突变导致 FTDP 17通过损害E10选择性剪接或tau功能。一个令人费解 FTDP 17综合征的一个方面是，不同的tau突变损害选择的 神经元和神经胶质的亚型。我们的第一个假设是， 脆弱性可能反映了tau亚型的细胞类型特异性扰动， 导致不同的表型。为了验证这一假设，我们将确定mRNA 正常脑细胞亚群中tau亚型的表达谱。 虽然正常人脑中3R和4R tau蛋白的比例为1：1，但从未有过 在神经元和神经胶质的亚群中被确定。在目标1中，我们将 从对照组石蜡包埋的组织切片中显微解剖神经元和神经胶质 大脑，对提取的RNA进行线性扩增，然后进行定量扩增。 实时RTPCR以测量每种tau同种型mRNA的相对水平。在Aim中 2，我们将类似地研究FTDP中相同的神经元和神经胶质细胞群， 17个大脑这些数据与疾病表型的相关性将阐明 FTDP的机制1 7.由于FTDP 17突变在不同的人中产生不同的表型， 第二种假说提出， 与tau基因或蛋白质相互作用的第二个基因影响 FTDP 17表型表现在不同受影响的 家庭成员目的3通过检查差异来检验这一假设 候选基因（即那些参与剪接，RNA稳定性， tau功能，其他细胞过程）之间的影响和未受影响的FTDP 17 大脑区域和控制大脑使用定制的cDNA宏阵列。的 这些目标的完成将促进对FTDP 17和相关的理解， Tau蛋白病，并可能提供新的诊断和治疗的目标。