The regulation mechanism of shRNAspten to NR2B after neuron subjected to anoxia insult
- VernacularTitle:下调PTEN基因对缺氧损伤后海马神经元NR2B受体调控机制研究
- Author:
Lixue CHEN
- Publication Type:Journal Article
- Keywords:
PTEN;
shRNAs;
Fura 2-AM;
AlamarBlue;
Colorimetric assay
- From:
Journal of Chongqing Medical University
1986;0(04):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the regulation of PTEN with RNAi on NR2B after OGD.Methods:In the OGD model with anaerobic gas mixture and deoxygenated,glucose-free extracellular solution,shRNAspten-GFP and unrelated control plasmid PshGFP transfection were done respectively.The quantities of NR2B in the neuronal membrane were quantitatively measured with colorimetric assay,and the expression of NR2B mRNA were analyzed by RT-PCR.The neuronal activity was analyzed by AlamarBlue andintracellular free calcium concentration were measured with Fura-2 AM by laser confocal microscope(LCM).Results:shRNAspten-GFP plasmid can been expressed successfully in cultured neurons.The study demonstrated that there are numerous NR2B subunit expression on neuronal membrane by colorimetric assay(OPD)assay and RT-PCR.The expression and quantity of neuronal NR2B with shRNAspten-GFP treatment was significantly decrease after oxygen and glucose deprivation(OGD)and reperfusion(P
