Chemical heredity epilepsy persistent state disease animal model and construction method and application thereof

A technology of status epilepticus and animal models, which is applied in the field of chemical genetic status epilepticus disease animal models and its construction, which can solve the problems of neuron death, high mortality, and inability to clarify the origin of status epilepticus lesions

Inactive Publication Date: 2020-10-30
THE FIRST AFFILIATED HOSPITAL OF CHONGQING MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These models can well induce status epilepticus, but in practical applications it is easy to find that the success rate of these models is low and the mortality rate is high. The death of a large number of neurons caused by toxicity makes these model mice unable to be reused, and it is also impossible to conduct before and after control experiments on the same mouse

Method used

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  • Chemical heredity epilepsy persistent state disease animal model and construction method and application thereof
  • Chemical heredity epilepsy persistent state disease animal model and construction method and application thereof
  • Chemical heredity epilepsy persistent state disease animal model and construction method and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] A method for constructing an animal model of chemical genetic status epilepticus

[0048] 1. Experimental animals

[0049] A total of 24 C57 mice aged 6-8 weeks were selected for this experiment, 12 each in Model 1 and Model 2. Among them, model 1 and model 2 are independent of each other, and model 1 induces status epilepticus by simultaneously activating excitatory neurons in hippocampal CA1 area (1.5, 2.0, 1.4) and ventral anterior nucleus VA area (1.25, 0.94, 3.75) through chemical genetics In the second model, status epilepticus was induced by simultaneously activating excitatory neurons in the BLA region of the amygdala (3.0, 2.0, 4.6) and the VA region of the ventral anterior nucleus of the thalamus (1.25, 0.94, 3.75).

[0050] 2. Experimental method

[0051] 2.1 Virus injection

[0052] 2.1.1 Routine treatment before operation

[0053] C57 mice were anesthetized by intraperitoneal injection of 1% pentobarbital sodium and fixed on a brain stereotaxic device (...

Embodiment 2

[0072] Comparison of chemical genetic status epilepticus disease animal model and Pilocarpine status epilepticus animal model

[0073] 1. Experimental animals

[0074] In this experiment, a total of 36 C57 mice aged 6-8 weeks were selected and divided into 3 groups with 12 mice in each group. The chemical genetic status epilepticus disease animal model in step 2.5 in Example 1 was selected.

[0075] 2. Experimental method

[0076] The construction of the animal model of Pilocarpine status epilepticus: commonly used Pilocarpine doses: 280mg / kg, 300mg / kg, 320mg / kg, 12 mice in each group, intraperitoneal injection of Pilocarpine to induce status epilepticus, and then statistics were made for each group Seizure duration within 30 minutes (min) from the onset of onset, and the final total model establishment success rate and mortality rate, the chemical genetic epilepsy constructed by the Pilocarpine epilepsy still needing state animal model and the condition of step 2.5 in Exampl...

Embodiment 3

[0084] Repeat induction experiment

[0085] 1. Experimental animals

[0086] The chemical genetic status epilepticus disease animal model constructed under the conditions of step 2.5 in Example 1, the Pilocarpine status epilepticus animal model constructed in Example 2

[0087] 2. Experimental method

[0088] With the chemical genetic status epilepticus disease animal model constructed under the conditions of step 2.5 in Example 1, with the Pilocarpine status epilepticus animal model constructed under the test conditions in Example 2, the same group of mice was subjected to the same test every other week. The induction experiment was repeated for a total of 4 experiments.

[0089] Table 3 Comparison of repeated induction duration between Pilocarpine animal model of status epilepticus and chemical genetic status epilepticus disease animal model

[0090]

[0091] Table 4 Comparison of repeated induction success rate and mortality between Pilocarpine animal model of status ...

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Abstract

The invention provides a chemical heredity epilepsy persistent state disease animal model and a construction method and application thereof. The epilepsy persistent state disease animal model is a mouse brain kernel group (a hippocampus CA1 region and a thalamus anterior nucleus VA region of a model I); injecting a brain stereotaxic virus (a chemical genetic virus rAAV-CaMKIIa-hM3D (Gq)-mCherry-WPREs-pA) in an apricot kernel BLA region and a thalamus anterior nucleus VA region on the outer side of a substrate of the model II, and embedding an electrode array in a mouse hippocampus CA3 region;after the mouse is recovered for one week, a metabolite CNO of clozapine is injected into the abdominal cavity so that the CNO is combined with a virus expression receptor, neurons are activated to induce epileptic persistent state attack, and the epilepsy persistent state disease animal model is obtained through behavioral observation and in-vivo multichannel local field potential recording judgment. The epilepsy persistent state disease animal model constructed by the method is stable in seizure duration, high in success rate, low in death rate and good in repeatability, and has important significance in researching the origin and formation mechanism of epilepsy persistent state, and screening and mechanism of drug-resistant epilepsy persistent state drugs.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a chemical genetic status epilepticus disease animal model and its construction method and application. Background technique [0002] Animal models currently used for the study of status epilepticus are almost all established using exogenous chemical poisons or electrical stimulation, which do not exist in humans or animals, and are not known causes of status epilepticus , the neurons involved in the activity, the nuclei of initiation and termination, and the location are also unclear in these animal models, so it is difficult for people to understand the "real world" of human status epilepticus through this animal model. [0003] The traditional animal model of status epilepticus mainly induces status epilepticus by intraperitoneal or intracranial injection of chemical drugs, which makes a large number of neurons overexcited and produces excitotoxicity. The most classic...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61D7/00A61N1/36
CPCA61D7/00A61N1/3605
Inventor 王学峰贺淼清
Owner THE FIRST AFFILIATED HOSPITAL OF CHONGQING MEDICAL UNIVERSITY
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