The changes of autophagic flux in primary cortical neurons after oxygen and glucose deprivation reoxygenation were detected
10.19405/j.cnki.issn1000-1492.2022.02.004
- Author:
Lei Zhang
1
,
2
;
Chao Dai
3
;
Yanfeng Han
4
;
Yang Li
1
;
Yuqi Hu
1
;
Fulei Chen
1
;
Dong Zhao
1
Author Information
1. Dept of Neurosurgery, The First Afiliated Hospital of Shihezi University, Shihezi 832000
2. Dept of Neurosurgery, Xinjiang Production and Construction Corps Tenth Division Beitun Hospital , Beitun 836000
3. Dept of Neurosurgery, Xinjiang Production and Construction Corps Tenth Division Beitun Hospital , Beitun 836000
4. Xinjiang Production and Construction Corps Tenth Division Disease Prevention and Control Center,Beitun 836000
- Publication Type:Journal Article
- Keywords:
cortical neuron cells;
autophagy;
autophagic flux;
OGD/R;
detect
- From:
Acta Universitatis Medicinalis Anhui
2022;57(2):182-186
- CountryChina
- Language:Chinese
-
Abstract:
Objective :To detect the changes of autophagic flux at different stages after oxygen⁃glucose deprivationreoxygenation (OGD/R) with several highly sensitive methods.
Methods :Primary cortical neurons after oxygen deprivation of sugar after reoxygenation (OGD/R) were divided into the experimental OGD/R group and OGD/R + bafilomycinA1 (BafA1) group , using an RFP⁃GFP series fluorescent tags LC3 gene transfection detection cytolysosome and fusion of lysosomes , transmission electron microscope (TEM) observation the ultrastructure of autophagy , p62/SQSTM1 combining LC3 protein to flip the experimental testing p62 and LC3 protein quantitative , p62 immune staining observing the distribution and content.
Results :Under fluorescence microscope , the ratio of autophagy lysosome to autophagosome increased significantly in OGD/R group , and the changes of autophagy structure in different stages could be observed in TEM. The ratio of soluble p62 and LC3 Ⅱ/ Ⅰ reflected the activation of autophagic
flux , and p62 was mainly distributed in BafA1 group after fluorescence staining.
Conclusion : Each method has its own advantages , and different methods and indicators can accurately reflect the specific changes of autophagic flux in different stages after neuronal OGD/R. Mastering and applying these methods can effectively explore central nervous system diseases from the perspective of autophagy.
- Full text:2024120510351639593精细化检测原代皮质神经元细...糖剥夺再复氧后自噬流的变化_张磊.pdf
