DEVELOPMENT OF CELLULAR AND ANIMAL MODELS FOR HUNTINGTONS DISEASE

亨廷顿病(HD)是一种常染色体显性遗传性神经退行性疾病 与CAG三核苷酸重复扩增相关的疾病。 HD患者(CAG)n三核苷酸重复序列多态范围为6~34 在正常个体中复制，但当重复序列扩大时是致病的 到35到126的范围。多聚谷氨酰胺的重复编码被定位 在348 kD HD蛋白的编码序列内。高清消息和 蛋白质被广泛表达，但没有解释其特定的 基底节的神经病理学。为了充分理解 HD的发病机制，我们正在进行以下具体的研究 目的：1)确定从沙门氏菌中分离纯化的HD蛋白的结构 杆状病毒表达系统，2)利用完整的 包含16、48和89个CAG重复序列的长度HD cDNA，3)至 利用含有HD的350 kb YAC建立YAC转基因小鼠 基因组基因座，并且已经被改造了48个重复，4)到 检查不同CAG重复大小对 HD-/-ES细胞的增殖、分化和凋亡 用含有不同CAG的HD表达载体转染 重复长度，以及5)检测转谷氨酰胺酶(TGase)的作用 通过共转染TGase表达载体在疾病过程中 与HD全长克隆到HD-/-ES细胞中。这些研究 将为HD蛋白的功能及其功能提供重要线索 在神经病理学、行为学和三核苷酸重复序列不稳定性中的作用 高清。

Huntington's Disease (HD) is an autosomal dominant, neurodegenerative disorder which is associated with CAG trinucleotide repeat expansions. The polymorphic (CAG)n trinucleotide repeat in HD ranges from 6 to 34 copies in normal individuals but is pathogenic when the repeats expand to a range of 35 to 126. This repeat coding for polyglutamines is located within the coding sequence of the 348 kD HD protein. The HD message and protein are widely expressed leaving unexplained its specific neuropathology in the basal ganglia. In order to fully understand the pathogenesis of HD, we are conducting studies with the following specific aims: 1) to determine the structure of the HD protein purified from a baculovirus expression system, 2) to develop transgenic mice using full length HD cDNA that contains either 16, 48 and 89 CAG repeats, 3) to develop YAC transgenic mice using a 350 kb YAC that contains the HD genomic locus and which has been retrofitted with 48 repeats, 4) to examine the effects of various CAG repeat sizes in the rate of proliferation, differentiation and apoptosis of HD-/- ES cells that have been transfected with HD expression constructs containing various CAG repeat lengths, and 5) to examine the effects of transglutaminase (TGase) in the disease process by co-transfecting a TGase expression construct with the HD full length clones into the HD-/- ES cells. These studies will provide important clues to the function of the HD protein, and its role in neuropathology, behavior and trinucleotide repeat instability in HD.