Method for constructing animal brain injury model based on hippocampal cholinergic neuron regulation and control

A cholinergic neuron and injury model technology, applied in the field of medicine, can solve the problems of lack of construction and evaluation of electromagnetic radiation brain injury animal models, lack of prevention and treatment measures, etc.

Active Publication Date: 2022-07-08
ACADEMY OF MILITARY MEDICAL SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003]Because the molecular mechanism of the electromagnetic radiation brain damage effect has not yet been elucidated, the target cell types and their molecular targets sensitive to electromagnetic radiation are also unclear, so far there is a lack of good

Method used

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  • Method for constructing animal brain injury model based on hippocampal cholinergic neuron regulation and control
  • Method for constructing animal brain injury model based on hippocampal cholinergic neuron regulation and control
  • Method for constructing animal brain injury model based on hippocampal cholinergic neuron regulation and control

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Embodiment 1

[0036] 1. Materials and methods

[0037] 1.1Chat-iCre mouse construction

[0038] Choline acetyltransferase (ChAT), a marker gene of cholinergic neurons, was used as the target gene, and P2A-iCre-P2A-tdTomato was inserted before the gene stop codon. The homology arms at the 5' and 3' ends are approximately 2 kb, respectively. Knock-in mice were prepared using the EGE system developed by Biositu based on CRISPR / Cas9. The schematic diagram of the targeting vector is as follows figure 1 shown.

[0039] 1.2Rosa26-ChR2 tool mouse construction

[0040] According to the characteristics of the mouse Rosa26 gene, CAG-loxP-Stop-loxP-ChR2-EGFP-WPRE-pA was inserted at the Rosa26 site. Gene knock-in mice were prepared using the EGE system developed by Biositu based on CRISPR / Cas9. The schematic diagram of the targeting vector is as follows figure 2 shown.

[0041] 1.3 Brain Stereotactic Injection of AAV and Fiber Optic Implantation

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Abstract

The invention discloses a method for constructing an animal brain injury model based on hippocampal cholinergic neuron regulation, which comprises the following steps: setting electromagnetic radiation conditions, and carrying out electromagnetic radiation treatment on a test animal; and comparing the influence of cholinergic neuron activation or inhibition of the hippocampal CA3 region of the experimental animal on the behavior of the experimental animal before and after the electromagnetic radiation treatment, and if the influence on the behavior of the experimental animal has statistical significance, constructing the electromagnetic radiation animal brain injury model by utilizing the electromagnetic radiation condition. According to the method, the electromagnetic radiation animal brain injury model can be well constructed, and possibility is increased for clarification of the molecular mechanism of the electromagnetic radiation brain injury effect and research and development of prevention and treatment drugs.

Description

technical field [0001] The present invention relates to the technical field of medicine. More specifically, the present invention relates to a method for constructing an animal brain injury model based on the regulation of hippocampal cholinergic neurons. Background technique [0002] Existing studies have found that electromagnetic radiation can cause changes in the morphological structure of brain tissue, including hippocampus, cerebral cortex, cerebellum, amygdala, and blood-brain barrier. Among them, the hippocampus is a key brain area involved in learning and memory functions. It is not only a long-term It is also involved in the formation of short-term memory and spatial memory, and is also involved in the regulation of emotional behavior. Our previous study found that the pathological changes of the hippocampus caused by microwave radiation were more obvious in the CA3 area. Using gene chip and proteomics technology, it was found that cytoskeleton damage, energy met...

Claims

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Application Information

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IPC IPC(8): A01K67/027
CPCA01K67/0275A01K67/0278A01K2227/105A01K2267/03A01K2217/206
Inventor 左红艳李杨郝延辉胡翠翠叶雨萌常晨旭孔晓旭鲁艳杰冯志华
Owner ACADEMY OF MILITARY MEDICAL SCI
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