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Method for constructing Alzheimer disease mouse model

An Alzheimer's disease and mouse model technology, applied in the field of biomedicine, can solve problems such as being unsuitable for large-scale drug screening, the model making cycle is too long, and the spread of tau pathology cannot be observed.

Active Publication Date: 2020-01-14
NANTONG UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] The second way is to select transgenic mice, such as 3XTg (Jackson laboratory mouse number 034830, genotype B6; 129-Psen1 tm1Mpm Tg(APPSwe,tauP301L)1Lfa / Mmjax), 6 weeks after injection of pathological tau extracted from the brain of AD patients at 12 months old, induced tau pathology can be detected; the disadvantage is that the model making cycle is still too long, more than one year Years, obviously not suitable for large-scale drug screening (Dai, C.L., Hu, W., Tung, Y.C., Liu, F., Gong, C.X., Iqbal, K., 2018. Tau passive immunization blocks seeding and spread of Alzheimer hyperphosphorylated Tau- induced pathology in 3x Tg-AD mice.Alzheimers ResTher 10,13.)
These mice developed tau pathology at 4.5 and 6 months of age, but because the transgene was expressed in neurons throughout the brain, the spread of tau pathology in hippocampal neurons to other neurons could not be observed later

Method used

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  • Method for constructing Alzheimer disease mouse model
  • Method for constructing Alzheimer disease mouse model
  • Method for constructing Alzheimer disease mouse model

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Example 1 Construction of Alzheimer's disease mouse model

[0023] Viral vectors integrating human tau protein truncations tau151-391, tau1-150, tau392-441 and tau protein were respectively constructed. The amino acid sequence of tau151-391 protein is shown in SEQ ID NO:1, and the DNA sequence is shown in SEQ ID NO:2; the amino acid sequence of tau1-150 protein is shown in SEQ ID NO:3, and the DNA sequence is shown in SEQ ID NO: 4; the tau392-441 protein amino acid sequence is shown in SEQ ID NO: 5, and the DNA sequence is shown in SEQ ID NO: 6; the tau protein amino acid sequence is shown in SEQ ID NO: 7, and the DNA sequence is shown in SEQ ID NO:8 shown.

[0024] The hSyn promoter-MCS-EGFP-3FLAG-SV40 PolyA virus vector was constructed, the full-length human tau protein and the corresponding DNA sequences of each truncated body were integrated, and the adeno-associated virus AAV9 was packaged.

[0025] According to the human tau protein truncated tau151-391, tau1-15...

Embodiment 2

[0065] Example 2 Pathological detection of rat brain tau

[0066] The sagittal 40 μM thick tissue sections of the brains of 15815 mice injected with different groups of viruses were washed with PBS, treated with PBS containing 0.5% Triton-X 100, and blocked with 5% goat serum blocking solution for 30 minutes, targeting phosphorylated tau (tau Pathology) specific antibody mouse-AT8 1:1000 was incubated overnight at 4 degrees, washed 3 times with PBS, Alexa-555 fluorescently labeled anti-mouse secondary antibody was incubated at room temperature for 2 hours in the dark, added 1:5000 Hochest for 15 minutes, washed three times with PBS, Patch cover. Fluorescence microscopy to detect tau pathology. The result is as Figure 2-4 As shown, the results show that 15815 female mice were injected with AAV9-tau151-391 in the brain at the age of 4 months, and at the age of 6 months, they were stained with AT8, a specific antibody for phosphorylated tau at the Ser202 / Thr205 site, to mark t...

Embodiment 3

[0069] Embodiment 3. The application example of mouse tau pathological model described in the present invention

[0070] (1) Screening genes that can inhibit tau pathology.

[0071] The target gene adeno-associated virus vector was constructed and injected into the hippocampus of 4-month-old 15815 mice to highly express the target gene. Two weeks later, AAV9-tau151-391 virus was injected at the same location to induce tau pathology. At the age of 6 months, the mouse brain was taken for immunofluorescence, and the intensity of tau pathology (the number of tau pathology-specific antibody mouse-AT8 positive cells) was compared between the mice injected with the target gene virus and the control virus, to determine whether the target gene affected the occurrence of tau pathology, In this way, genes that can effectively inhibit tau pathology are screened out.

[0072] (2) Screening drugs that can inhibit tau pathology

[0073] AAV9-tau151-391 virus was injected into the hippocam...

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Abstract

The invention discloses a method for constructing an Alzheimer disease mouse model. A genotype FVB-Fgf14Tg (tetO-MAPT* P301L) 4510 Kha / JlwsJ female mouse is adopted, a truncated tau protein or a vector expressing the truncated tau protein is injected into the brain of the mouse at the age of 4 months and then the mouse is fed for 2-4 months, and the amino acid sequence of the truncated tau proteinis as shown in SEQ ID NO: I. The Alzheimer disease mouse model is successfully constructed, tau pathology only appears in the hippocampus region of the constructed mouse model, and does not appear inother brain regions, the construction period is shortened to 6 months, and more than 50% of modeling time is saved.

Description

technical field [0001] The invention belongs to the field of biomedicine, and in particular relates to a method for constructing an Alzheimer's disease mouse model Background technique [0002] Alzheimer's Disease (AD for short), commonly known as senile dementia, is a common neurodegenerative disease in the elderly, manifested as learning and memory impairment, aphasia, apraxia, agnosia, and personality and behavior changes. dementia. AD has two major neuropathological features: extracellular amyloid plaques (amyloid plaques) formed by the accumulation of β-amyloid (Aβ) and intracellular neuronal neurons mainly formed by abnormally hyperphosphorylated tau protein. Fibrous tangle (Neurofibrillary tangle, NFT). So far, all clinical trials targeting Aβ have shown no significant effect (Iqbal, K., Liu, F., Gong, C.X., 2018. Recent developments with tau-based drug discovery. Expert Opin Drug Discov 13, 399-410.). More and more studies suggest that the spread of tau pathology ...

Claims

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Application Information

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IPC IPC(8): A01K67/027
CPCA01K67/0278
Inventor 储丹丹周艳王渺得周鼎伟吴若桢
Owner NANTONG UNIVERSITY
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