TRANSOMIC MOUSE MODEL FOR ALZHEIMER'S DISEASE

阿尔茨海默病(AD)是一种常见的痴呆，具有以下特点 神经病理学。尽管已经提出了许多动物模型 它们表现出一些神经化学和病理异常 在AD中，没有模型可以用来明确地研究 该病的发病机制和治疗。我们建议开发一种 选择染色体区域的AD转基因小鼠模型，认为 含有致病基因(S)的基因，是从患有 家族性阿尔茨海默病(FAD)并插入到 受体小鼠(“转基因小鼠”)。我们开发的方法是为了 精确显微解剖染色体条带并显微注射被解剖的 基因组DNA，在其自然环境中，进入受精的小鼠卵子。这个 从组织培养细胞的染色体中显微切割的区域有 21q11-q22在早发性FAD(伴有和不伴有淀粉样蛋白)患者中的作用 前体蛋白(APP)717突变)和19q13基因突变 开始流行。将进行实验，其中解剖的碎片来自 21号染色体将同时包含推测的FAD基因和APP基因，如 以及FAD基因和APP基因的解剖实验 分别显微注射到小鼠胚胎中。相同的地区 将从正常人身上解剖出来，作为基因剂量的对照。 21q11-q22也将从遗传性荷兰患者身上解剖出来。 脑出血合并淀粉样变性(HCHA-D)是否存在不同的APP 突变会导致转基因小鼠出现不同的表型。出生的活产自 将使用聚合酶链式反应方法对感染的胚胎进行人类序列筛选 检测人类特定的Alu序列以及杂合子和纯合子 通过选择性交配培育出的易体小鼠品系。我的大脑 转基因小鼠将接受详细的神经病理检查， 尤其是老年斑和神经原纤维缠结，以及 星形胶质细胞增多症和β淀粉样蛋白沉积的免疫组织化学。脱氧核糖核酸 来自表现出神经病理的转体小鼠品系的研究将用 关于特定人类序列的组织和表达 (特别是APP基因)也使用了过滤杂交技术 作为确定人类序列在转体小鼠中的位置 基因组原位杂交技术。总而言之，这种方法具有 建立合法的阿尔茨海默病小鼠模型的可能性 明确识别人类染色体图谱中导致 FAD中的特定神经病理。

Alzheimer's disease (AD) is a common dementia with characteristic neuropathology. Although a number of animal models have been proposed which exhibit some of the neurochemical and pathological abnormalities seen in AD, there is no model which can be utilized to definitively study the pathogenesis and treatment of this disease. We propose to develop a transgenic mouse model of AD in which selected chromosomal regions, thought to contain the disease-causing gene(s), are dissected from patients with familial Alzheimer's disease (FAD) and inserted into the germ line of recipient mice ("transomic mice"). We have developed the methodology to precisely microdissect chromosome bands and microinject the dissected genomic DNA, in its natural context, into fertilized mouse ova. The regions to be microdissected from chromosomes from tissue culture cells are 21q11-q22 in patients with early onset FAD (with and without an amyloid precursor protein (APP) 717 mutation) and 19q13 from patients with late onset FAD. Experiments will be performed in which dissected fragments from chromosome 21 will contain both the putative FAD locus and the APP gene, as well as experiments in which the FAD locus and APP gene are dissected separately and microinjected into mouse embryos. The identical regions will be dissected from normal individuals as a control for gene dosage. 21q11-q22 will also be dissected from Dutch patients with hereditary cerebral hemorrhage with amyloidosis (HCHA-D) to determine if different APP mutations induce varying phenotypes in transgenic mice. Live births from infected embryos will be screened for human sequences using a PCR method to detect human specific Alu sequences, and heterozygous and homozygous transomic mouse strains developed by selective matings. Brains of transomic mice will be examined in detail for neuropathologic lesions, particularly senile plaques and neurofibrillary tangles, and by immunohistochemistry for astrocytosis and beta amyloid deposition. DNA from transomic mouse strains exhibiting neuropathology will be studied with respect to the organization and expression of specific human sequences (particularly the APP gene) using filter hybridization techniques as well as determining the location of human sequences in the transomic mouse genome using in situ hybridization. In summary, this methodology has the potential of establishing a legitimate mouse model for AD and to unambiguously identify regions of the human chromosome map which cause specific neuropathology in FAD.