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Mouse model for studying axon regeneration of spinal anterior horn motor neurons

A technology of motor neuron and mouse models, applied in the field of animal models, can solve problems such as research on axon regeneration of neurons that cannot be used

Inactive Publication Date: 2019-04-16
SUZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It is limited in that it cannot be used to study the regeneration of motor neuron axons located in the anterior horn of the spinal cord

Method used

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  • Mouse model for studying axon regeneration of spinal anterior horn motor neurons
  • Mouse model for studying axon regeneration of spinal anterior horn motor neurons
  • Mouse model for studying axon regeneration of spinal anterior horn motor neurons

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1: Localized injection of AAV-cre virus in the anterior horn of the spinal cord

[0034] use figure 1 According to the method, the mouse spinal cord was exposed, and then the AAV-cre virus was injected into the anterior horn of the mouse spinal cord according to the following method:

[0035] 1. Needle for injection: The material used is PCR microinjection glass tube (Drummond, #5-000-1001-X10), and the glass tube is pulled into a needle for injection with a needle puller (NARISHIGE, PC-10).

[0036] 2. Virus: virus titer is 1×10 13 , the injection volume is 1ul.

[0037] 3. Injection equipment: Brain Stereotaxic Apparatus (Shenzhen Ruiwode Company).

[0038] 4. Model mice: Rosa-tdTomato flox conditional gene knockout mice.

[0039] 5. Injection steps: install the microinjection needle with 1ul AAV-cre virus on the brain stereotaxic apparatus, and inject AAV-Cre virus at the anterior angle position of mouse spine L1 ( figure 2 ), the injection coordinates ...

Embodiment 2

[0041] Neurons infected with AAV-cre virus express Tomato red fluorescent protein. First, we observed the expression of Tomato protein at the injection site of the anterior horn of the spinal cord, and then used ChAT-specific antibodies to label motor neurons to detect whether motor neurons were labeled. The axons from the motor neurons in the anterior horn of the spinal cord transmit to the sciatic nerve through the dorsal root ganglion (DRG), and finally extend to the motor endplate of the muscle. Therefore, we took the spinal cord, DRG, sciatic nerve and muscle to detect the expression of Tomato red fluorescent protein to confirm that the virus successfully infected the motor neurons in the anterior horn of the spinal cord.

[0042] (1) Detection of the expression of Tomato protein in the anterior horn of the spinal cord. Two weeks after injecting AAV-cre virus into the anterior horn of the spinal cord of Rosa-tdTomato mice, the tissue was taken for frozen section, and the...

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Abstract

The invention discloses a mouse model for studying axon regeneration of spinal anterior horn motor neurons, and belongs to the technical field of animal models. The mouse model is built for the firsttime by using AAV-cre virus and Rosa-tdTomato flox conditional gene knockout mice, and a method and coordinates for stereotactic injection in the spinal anterior horn are provided, marking the spinalanterior horn motor neurons. The built mouse model is provided for studying axon regeneration of the spinal anterior horn motor neurons.

Description

technical field [0001] The invention relates to a mouse model for studying the axon regeneration of the anterior horn motor neuron of the spinal cord, and belongs to the technical field of animal models. Background technique [0002] With the rapid development of modern transportation, sports activities, and construction engineering, nerve injuries, especially spinal cord injury (SCI), are increasing day by day. The spinal cord is an important part of the central nervous system (CNS), and its main function is to transmit nerve information between the brain and peripheral nerves; at the same time, it is also the low-level center of many reflex activities. Severe spinal cord injuries often result in permanent loss of sensory, motor, and sphincter functions below the level of injury, leading to paralysis. Its high disability rate and high medical expenses have brought a heavy burden to the patient's family and even the whole society. According to statistics, the incidence of ...

Claims

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Application Information

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IPC IPC(8): A61K35/76
CPCA61K35/76
Inventor 赛吉拉夫马艳霞王丰张鸿程齐士斌马进进张浩楠徐金辉秦绪祯
Owner SUZHOU UNIV
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