Tau Isoform Expression in FTDP-17

FTDP-17 中的 Tau 同工型表达

• 批准号: 6540426
• 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/6540426
• 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.

描述（由申请人提供）： 高磷酸化的tau是额颞的标志性脑病变 痴呆症与帕金森氏症与染色体17（FTDP 17）有关 由不同的tau基因突变引起的带有多种表型的tauopathy。 tau 是微管（MT）结合蛋白，可促进微管蛋白聚合到 MTS并稳定MT。成年人类大脑包含六个TAU同工型，一半 具有3个3R同工型）和一半，有4个C端MTBIND重复序列（4R同工型） 通过外显子10（E 10）的剪接产生。 tau基因突变引起 FTDP 17通过损害E 10替代剪接或TAU功能。令人困惑 FTDP 17综合征的方面是选择了不同的tau突变损害 神经元和神经胶质的亚型。我们的第一个假设是这种选择性 脆弱性可能反映了细胞类型的特异性tau同工型的扰动 导致各种表型。为了检验该假设，我们将确定mRNA 正常脑细胞亚群中Tau同工型的表达谱。 尽管在正常人大脑中，3R与4R tau的比率为1：1，但它从来没有 在神经元和神经胶质的亚群中确定。在AIM 1中，我们将 来自控制的组织切片的微解答神经元和神经胶质 大脑，对提取的RNA进行线性扩增，然后进行定量 实时RTPCR测量每个TAU同工型mRNA的相对水平。目标 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澄清 FTDP1 7的机制。由于FTDP1 7突变在中产生不同的表型 相同的亲属，第二个假设提出了改变表达的表达 与tau基因或蛋白质相互作用的第二个基因会影响 FTDP 17的表型表现形式在不同受影响的 家庭成员。 AIM 3通过检查差异来检验该假设 候选基因的表达（即参与剪接，RNA稳定性的基因， tau功能，其他细胞过程）在受影响和未受影响的FTDP 17之间 大脑区域和控制大脑使用Custommade cDNA宏观阵列。这 这些目标的完成将提高人们对FTDP 17的了解 tauopathies，可能为诊断和治疗学提供新的靶标。