Method for preparing midbrain dopaminergic neuron

Abstract

The invention discloses a method for building PITX3/GFP/A53T alpha-synuclein three-genotype mice. The method comprises the following steps of (1) respectively building PITX3-IRES2-tTA, tetO-A53T alpha-synuclein and tetO-H2Bj/GFP transgenic mice; (2) performing hybridization on the PITX3-IRES2-tTA transgenic mice in the step (1) with one of the other two kinds of transgenic mice to obtain double-genotype mice; (3) performing hybridization on the double-genotype mice with the third kind of transgenic mice in the step (1) to obtain three-genotype mice. The invention also discloses a method for preparing midbrain dopaminergic neuron. The method comprises the following steps of building PITX3/GFP/A53T alpha-synuclein three-genotype mice and separating the midbrain dopaminergic neuron from the three-genotype mice according to the GFP expression. The method provided by the invention uses GFP as report genes; the physiologic properties and functions of the midbrain dopaminergic neuron are notinfluenced; meanwhile, the expression position of the alpha-synuclein gene in the midbrain dopaminergic neuron can be marked, so that the high-purity midbrain dopaminergic neuron with the GFP fluorescence labeling can be specifically separated out.

Description

technical field [0001] The invention belongs to the field of neurobiology, and in particular relates to a method for preparing midbrain dopamine neurons. Background technique [0002] Parkinson's disease (Parkinson's Disease, PD) is the second most common degenerative disease of the central nervous system. Lewy neurite-like structures. The etiology of dopamine neuron reduction is complex, including genetic factors, post-genetic factors, and environmental risk factors. In the past fifteen years, genetic studies of rare familial PD have identified single-gene mutations, including mutations in the dominant genes α-synuclein and LRRK2, and recessive genes parkin, DJ-1, and PINK1 In recent years, the results of genome-wide research have further confirmed that the mutation of genetic factors is the main cause of common sporadic PD; the above genetic studies have proved that genetic factors play a key role in the occurrence and development of PD. α-synuclein is the main componen...

AI Technical Summary

Problems solved by technology

Limited to the current level of science and technology, it is very difficult for scientists to accurately isolate dopamine neurons in the midbrain. At present, there is no method for accurately isolating dopamine neurons from midbrain tissue, which greatly limits the ability of α-synuclein gene. Study on the degeneration mechanism of midbrain dopamine neurons

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