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A method for constructing a live animal model for studying neuronal autophagy

A model and genotype technology, applied in animal husbandry and other directions, can solve the problem of little understanding of the exact mechanism

Active Publication Date: 2019-06-28
SHANGHAI INST OF ORGANIC CHEM CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Although there is some evidence that autophagy is important for axonal as well as neuronal homeostasis, its precise mechanisms are poorly understood

Method used

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  • A method for constructing a live animal model for studying neuronal autophagy
  • A method for constructing a live animal model for studying neuronal autophagy
  • A method for constructing a live animal model for studying neuronal autophagy

Examples

Experimental program
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Embodiment 1

[0067] Embodiment 1, the construction of the animal model about neuronal autophagy

[0068] First, the genotype dpr-Gal4 (chromosome II) (derived from Bloomington stock) was crossed with the genotype UAS-mCD8-GFP (chromosome II) (derived from Bloomington stock), and then the offspring I was selected to contain this gene at the same time. Two transgenic dpr-Gal4 / UAS-mCD8-GFP virgin fruit flies were crossed with yw (chromosome I) (from Bloomington stock) fruit flies, and the progeny fruit flies were crossed The male fruit fly with the darkest eye color among the flies is selected (if there are fruit flies with white eyes in the offspring, it means that homologous recombination has occurred), and then the male fruit fly with darker eye color is selected and the female fruit fly genotype is yw The virgin flies of Sco / Cyo (derived from A.Sehgal) were hybridized until the larvae of the third generation of the child appeared, and the corresponding male fruit flies of the second gener...

Embodiment 2

[0069] Example 2. Study on the changes of autophagy under different conditions of axon injury

[0070] The Drosophila wing model is used to study the changes of autophagy at different time points of injury. The specific embodiment is as follows: figure 2Shown: the first longitudinal row is the control group, that is, the Drosophila wing is not damaged; the second and third longitudinal row are the experimental groups, and the time points of nerve bundle damage are different. In the control group, the mcherry fluorescence signal was relatively weak and distributed scatteredly on the axons marked by mCD8-GFP when the wing of Drosophila was not damaged. And after 0.5 hours of axonal injury (such as figure 2 shown), the fluorescent signal of mcherry-Atg8a on the axons is also lower than that of the uninjured axons, and the axons marked by mCD8-GFP at this time are relatively intact; after 6 hours of injury, there is an obvious The level of autophagy increases, while the axon i...

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Abstract

The invention provides a method for establishing a live animal model for studying neuron autophagy. According to the method, the wing nerve tract of a live fruit fly is labeled with fluorescin, an autophagy-related gene is labeled with different fluorescin at the same time, and then the change of autophagy on the axon under the condition of wing damage handling or fruit fly drug feeding can be rapidly and clearly observed. The experimental result shows that neuron autophagy can be simply and directly studied with the animal model established through the method.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular, the invention relates to a method for constructing a living animal model for studying neuron autophagy. Background technique [0002] Autophagy is an organism's self-digestion mechanism that removes damaged organelles and non-functional proteins in cells through lysosomes. Autophagy is divided into three categories according to the mechanism of substrate delivery to lysosomes: macroautophagy (Macroautophagy), microautophagy (Microautophagy) and chaperone-mediated autophagy (CMA). [0003] Macroautophagy is the main autophagy pathway, and it is also the most studied type of autophagy. In the process of macroautophagy, a double-membrane autophagosome (Autophagosome) will be formed, and the protein products of some autophagy-related genes (Autophagy-related genes, Atg) conserved in evolution participate in the regulation of this pathway. Under the induction of factors such as starvation...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01K67/033
CPCA01K67/033
Inventor 方燕姗王海琼曹旭王钊
Owner SHANGHAI INST OF ORGANIC CHEM CHINESE ACAD OF SCI
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