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Mouse neurotrophic keratitis animal model and application thereof

A mouse model and neurotrophic technology, which is applied in animal/human peptides, compound screening/testing, biochemical equipment and methods, etc., can solve problems such as long operation time, varying degrees of injury, and poor model consistency, and achieve reliable results. Highly repeatable and model-stable effects

Pending Publication Date: 2021-01-22
QINGDAO UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] At present, the mouse model of trigeminal nerve ophthalmic branch injury is commonly used. This method involves craniotomy and damages the cranial brain. The brain damage of the animal is large, and there are many complications (such as changes in intracranial pressure). Sutures are required after surgery, and the wound Long healing time, cumbersome postoperative care procedures, high probability of wound infection, long operation time, poor model consistency, and varying degrees of injury

Method used

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  • Mouse neurotrophic keratitis animal model and application thereof
  • Mouse neurotrophic keratitis animal model and application thereof
  • Mouse neurotrophic keratitis animal model and application thereof

Examples

Experimental program
Comparison scheme
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Embodiment 1

[0028] Example 1: Construction of AQP5-KO mouse NK animal model

[0029] 1. Construction and identification of AQP5-KO mice

[0030] AQP5 (NCBI ID: 11830) is located on chromosome 15 of the mouse. For the information of AQP5-KO mice and the knockout region, see figure 1 A, B, AQP5-KO mice were obtained by knocking out the chr15:99589876-99594444 gene fragment with a length of 4569 bp by using CRISPR / Cas9 technology and applying high-throughput electroporation to fertilized eggs.

[0031] figure 1 C is the sequencing identification result of the AQP5-KO mouse, compared with the wild-type sequence, it can be seen that a total of 4569bp sequence is missing, indicating that the model mouse was successfully established.

[0032] In subsequent routine feeding, PCR amplification was used to detect the knockout effect of AQP5-KO mice.

[0033] PCR amplification target fragment: reaction conditions and reaction system:

[0034] (1) PCR reaction conditions: 94°C for 3min; 94°C for 3...

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Abstract

The invention provides a neurotrophic keratitis (NK) mouse model, namely an AQP5-KO model mouse. The mouse model is constructed by reducing the expression quantity of a gene AQP5 or knocking out the gene AQP5. The AQP5-KO model mouse disclosed by the invention is high in repeatability and stable in model, is beneficial to research and drug intervention of a dry eye pathomechanism, and can avoid the influence of other influence factors on an experimental result. After the AQP5-KO model mouse is born, the characteristic change of corneal subepithelial nerve reduction occurs; and after birth, thecorneal sensitivity is obviously reduced, and the corneal sensitivity is not obviously changed along with increase of age.

Description

technical field [0001] The invention belongs to the technical field of medical animal models, and in particular relates to a method for establishing an animal model of neurotrophic keratitis. Background technique [0002] As an avascular tissue, the cornea is the most densely innervated structure in the human body. Corneal nerves are responsible for maintaining the structural and functional integrity of the cornea, conveying sensations of touch, temperature, and pain, and playing a role in the blink reflex, wound healing, and tear secretion. Corneal nerve damage leads to corneal sensory loss and nutritional disorders, which can cause degenerative keratopathy, including corneal epithelial shedding, persistent corneal epithelial defect, corneal ulcer, matrix dissolution, corneal perforation and other different clinical manifestations. [0003] Neurotrophic keratitis (NK) is a degenerative corneal disease caused by damage to the function of the corneal nerve innervated by the ...

Claims

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Application Information

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IPC IPC(8): C12N15/87A01K67/027A61K49/00C12N15/11C12Q1/6888
CPCC12N15/87A01K67/0276C07K14/47A61K49/0008C12Q1/6888A01K2267/03A01K2227/105
Inventor 陈鹏狄国虎陈豪
Owner QINGDAO UNIV
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