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的神经退行性疾病， 与CAG三核苷酸重复扩增相关的疾病。 HD的CAG n三核苷酸重复序列多态性范围为6 ~ 34 在正常个体中复制，但当重复序列扩展时是致病的 范围为35至126。多聚谷氨酰胺的重复编码位于 在348 kD HD蛋白的编码序列内。HD消息和 蛋白质被广泛表达， 基底神经节的神经病理学为全面了解 HD的发病机制，我们正在进行以下具体的研究 目的：1）确定从大肠杆菌中纯化的HD蛋白的结构， 杆状病毒表达系统，2）使用完整的杆状病毒表达系统开发转基因小鼠， 长度HD cDNA，其含有16、48和89个CAG重复序列，3）至 使用含有HD的350 kb YAC开发YAC转基因小鼠 基因组位点，并已被改造为48个重复序列，4）， 检查各种CAG重复序列大小对 HD-/- ES细胞的增殖、分化和凋亡， 用含有各种CAG的HD表达构建体转染 重复长度，和5）检查转氨酶（TGase）的影响， 通过共转染TGase表达构建体， 将HD全长克隆导入HD-/- ES细胞。这些研究 将为HD蛋白的功能提供重要线索， 在神经病理学、行为学和三核苷酸重复不稳定性中的作用 HD.

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.