Affiliation:
1. the First Affiliated Hospital of Zhengzhou University
Abstract
Abstract
Objectives
To investigate the expression of adaptor protein complex-4 and GluR2 in oxygen-glucose deprived neurons, and to investigate the relation between AP-4 and GluR2 in injured neurons.
Methods
We used primary cultured hippocampal neurons from 18-day-old Sprague-Dawley rat fetuses as material. Oxygen-glucose deprivation was performed to build a cellular model of hypoxic-ischaemic brain injury. Real-time RT-PCR and Western blotting were performed to evaluate the expression of GluR1, GluR2 and AP4M1, both in mRNA and protein levels. AP4M1 and GluR1/GluR2 were double-labeled using immunofluorescence to investigate whether their distributions were similar. Finally, we performed co-immunoprecipitation after over-expressing AP4M1 to investigate whether AP4M1 interacts with GluR2.
Results
We found that AP4M1 was down-regulated in oxygen-glucose-deprived neurons, while the levels of GluR1 and GluR2 were not reduced. After the OGD procedure, both AP4M1 and GluR2 showed similar distributional pattern. This phenomenon was not observed in GluR1 distribution. AP4M1 cannot be co-immunoprecipited with GluR2 with normal AP4M1 expression, but the CO-IP between AP4M1 and GluR2 turned to positive when AP4M1 is over-expressed.
Conclusion
AP4M1 mediates GluR2 trafficking in oxygen-glucose deprived neurons, either directly or through other proteins.
Publisher
Research Square Platform LLC
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