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摘要：

蓝斑神经元的死亡数目与β淀粉样蛋白沉积及阿尔茨海默病严重程度成正相关。携带有突变β-淀粉样前体蛋白基因和突变早老素1基因APP/PS1双转基因小鼠阿尔茨海默病模型是目前研究阿尔茨海默病较好的动物模型，但围绕该模型的研究主要关注海马和皮质改变。鉴于此，实验选用五六月龄年轻组雌性APP/PS1双转基因小鼠与及十六七月龄老年组APP/PS1双转基因小鼠进行实验，每组均设同龄野生型小鼠作对照。免疫组织化学染色结果显示，与老年组野生型小鼠比较，老年组APP/PS1双转基因小鼠蓝斑儿茶酚胺能神经元特异性标志物-酪氨酸羟化酶阳性神经元胞体变大，细胞总数量减少23%，酪氨酸羟化酶阳性纤维少而粗短、有断裂。无偏差体视学方法定量分析结果显示，老年组APP/PS1双转基因小鼠蓝斑区酪氨酸羟化酶阳性神经元平均体积较年轻组增加，并与蓝斑总体积呈正相关。说明APP/PS1双转基因小鼠老年小鼠蓝斑去甲肾上腺素神经元和纤维较易发生退行性病理改变。

关键词:

Abstract:

Mice carrying mutant amyloid-β precursor protein and presenilin-1 genes (APP/PS1 double trans-genic mice) have frequently been used in studies of Alzheimer’s disease; however, such studies have focused mainly on hippocampal and cortical changes. The severity of Alzheimer’s disease is known to correlate with the amount of amyloid-β protein deposition and the number of dead neurons in the locus coeruleus. In the present study, we assigned APP/PS1 double transgenic mice to two groups according to age: young mice (5–6 months old) and aged mice (16–17 months old). Age-matched wild-type mice were used as controls. Immunohistochemistry for tyrosine hydroxylase (a marker of catecholaminergic neurons in the locus coeruleus) revealed that APP/PS1 mice had 23% fewer cells in the locus coeruleus compared with aged wild-type mice. APP/PS1 mice also had increased numbers of cell bodies of neurons positive for tyrosine hydroxylase, but fewer tyrosine hydroxylase-positive fibers, which were also short, thick and broken. Quantitative analysis using unbiased stereology showed a significant age-related increase in the mean volume of tyrosine hy-droxylase-positive neurons in aged APP/PS1 mice compared with young APP/PS1 mice. Moreover, the mean volume of tyrosine hydroxylase-positive neurons was positively correlated with the total volume of the locus coeruleus. These findings indicate that noradrenergic neurons and fibers in the locus coeruleus are predisposed to degenerative alterations in APP/PS1 double transgenic mice.

对双转基因APP/PS1小鼠动物进行研究

双转基因APP/PS1小鼠AD转基因app/ps1小鼠制备构建方法：

双转基因APP/PS1小鼠AD转基因app/ps1小鼠品系描述：

双转基因APP/PS1小鼠AD转基因app/ps1小鼠可表达突变的人类早老素（DeltaE9）和人鼠淀粉样前蛋白（APPswe）融合体，APP/PS1双转基因小鼠AD转基因app/ps1小鼠这两个基因的表达都由小鼠朊病毒蛋白启动子启动。人类早老素基因的DeltaE9突变是该基因的第九个外显子缺失产生的，此突变会导致早发性老年痴呆症。对APP/PS1双转基因小鼠AD转基因app/ps1小鼠脑蛋白匀浆进行免疫检测发现，人类早老素蛋白高水平地替代了可检测到的APP/PS1双转基因小鼠AD转基因app/ps1小鼠内源性蛋白，并且，在APP/PS1双转基因小鼠AD转基因app/ps1小鼠脑匀浆中还检测到了人源淀粉样前蛋白。据研究者报道，6-7月龄的APP/PS1双转基因小鼠AD转基因app/ps1小鼠脑内会形成beta淀粉状蛋白沉淀。

双转基因APP/PS1小鼠AD转基因app/ps1小鼠制备引物应用领域

1) 老年痴呆症的人/鼠基因同系物研究

2) 老年痴呆症和神经退行性变化的神经生物学研究

目的对所获的APP/PS1双转基因小鼠AD转基因app/ps1小鼠阿尔茨海默病(A lzhe im er d isease,AD)模型小鼠进行基因鉴定,进一步对其进行组织学分析,检测老年斑(sen ile p laque,SP)的形成情况。方法设计特定的引物,PCR扩增转入基因组DNA中的APP基因,对转入APP/PS1双转基因小鼠老年痴呆AD小鼠应用刚果红染色结合免疫组化观察Aβ沉积、小胶质细胞和星型胶质细胞的活化。结果与未转入的阴性对照相比,在AD的APP/PS1双转基因小鼠皮质和海马内可见Aβ斑块形成,围绕在Aβ斑块周围的小胶质细胞和星形胶质细胞处于活化状态,形成典型的SP结构。结论我们获得的APP/PS1双转基因小鼠老年痴呆AD小鼠能够模拟AD患者脑内的

STRAIN NAME: C57BL/6J--KO（Apoe-KOXCMV-hApoE4）
TYPE: transfergen
BACKGROUND STRAIN: C57BL/6J
COAT COLOR: black
ORIGIN: Developed through crosses and back-crosses between CMV-ApoE4 transgenic mice and ApoE4 gene knockout mice. Systemic expression human ApoE4 gene, but no expression mice ApoE3 gene, IT is important tool to study different ApoE alleles. The human ApoE CDNA length 1.163kb. ApoE gene has three alleles, namely E2, E3 and E4, constitute the six different genotypes: three homozygous (E2/E2, E3/E3 and E4/E4) and three heterozygous (E2 / E3, E3/E4 and E2/E4). The gene frequencies is differen in differen ethnic and different regional, E2 accounted for 8%, E3 accounted for 78%, E4, 14%. using the CMV expression ApoE2, ApoE3 and ApoE4 transgenic mice is mportant models for study CHD, hyperlipidemia, cerebral infarction and chronic hepatitis and other diseases

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