Endogenous epileptic seizure animal model and construction method thereof

Abstract

The invention discloses an endogenous epileptic seizure animal model. The endogenous epileptic seizure animal model is constructed by adopting chemogenetics or optogenetics activation virus to be injected to a hippocampus CA3 region of a mouse and expressed for three weeks, wherein the activation virus is selected from: (a) a mixture of rAAV-CaMKIIa-hM3D(Gq)-mCherry-WPREs-pA and rAAV-VGAT1-hM3D(Gq)-mCherry-WPRE-pA, and (b) a mixture of rAAV-Ef1[alpha]-DIO-hChR2(H134R)-EYFP-WPRE-pA and rAAV-CaMKII-CRE-WPRE-pA as well as AAV-VGAT1-CRE-WPRE-pA. Compared with a CA1 region and VA region multi-point injection model, the epileptic seizure animal model is more suitable for mechanism research of epilepsy and screening of antiepileptic drugs.

Description

technical field [0001] The invention belongs to the field of medicine, and specifically relates to an endogenous epileptic seizure animal model and a construction method thereof, in particular to an endogenous epileptic seizure animal model constructed by chemical genetics or optogenetics activation virus and a construction method thereof. Background technique [0002] Refractory epilepsy has not made significant progress in the past 40 years. The reason is that the pathogenesis is not fully understood. Previous studies on the pathogenesis of epilepsy were mostly based on traditional animal models of epilepsy, such as pilocarpine, kainic acid, or electrical stimulation. These exogenous animal models of epilepsy are very different from human epilepsy. The brain regions involved or the specific neurons involved in the seizure process are also unclear. Therefore, these exogenous epilepsy models cannot yet fully mimic human epilepsy. The current view is that the occurrence of...

AI Technical Summary

Problems solved by technology

First, for example, pilocarpine, kainic acid, and PTZ models, using diffusion chemicals that are inherently variable, do not precisely pinpoint diseased areas and cell types, and make it difficult to distinguish the contribution of plasticity mechanisms to tissue damage

Second, the experimenter has no control over the subset of cells activated upon ignition, making it difficult to establish a causal relationship between cell types and pathological outcomes

There are still many deficiencies in the construction method of the model

First, this method needs to inject virus at multiple points in the brain (such as CA1 area, VA area, BLA area, etc.) to establish a status epilepticus model, but two or more nuclei are activated at the same time, and it is difficult to identify nuclei at the same time. Upstream and downstream relationships, significantly limited in epilepsy network studies

Second, the model established by injecting viruses at multiple points in the brain is not conducive to the study of the evolution of neurotransmitters in the neural network during epileptic seizures

Third, the time and cost of multi-point injection of virus in the same mouse brain is relatively long, and the amount of virus is relatively large

If the upstream and downstream virus injections are required to be injected at the same time, it is easy to cause the death of the mice during the injection process; if the injections are separated, the experiment time will be extended and the time cost will be increased.

At the same time, it also increases the expense and cost of mouse feeding

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