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. 2012 Jan 25;28(1):69–76. doi: 10.1007/s12264-012-1065-5

Knock-down of postsynaptic density protein 95 expression by antisense oligonucleotides protects against apoptosis-like cell death induced by oxygen-glucose deprivation in vitro

Jing-Zhi Yan 1,2, Yong Liu 1,2, Yan-Yan Zong 1,2, Guang-Yi Zhang 1,2,✉
PMCID: PMC5560291  PMID: 22233891

Abstract

Objective

Postsynaptic density protein 95 (PSD-95) plays important roles in the regulation of glutamate signaling, such as that of N-methyl-D-aspartate receptors (NMDARs). In this study, the functional roles of PSD-95 in tyrosine phosphorylation of NMDAR subunit 2A (NR2A) and in apoptosis-like cell death induced by oxygen-glucose deprivation (OGD) in cultured rat cortical neurons were investigated.

Methods

We used immunoprecipitation and immunoblotting to detect PSD-95 protein level, tyrosine phosphorylation level of NR2A, and the interaction between PSD-95 and NR2A or Src. Apoptosis-like cells were observed by 4,6-diamidino-2-phenylindole staining.

Results

Tyrosine phosphorylation of NR2A and apoptosis-like cell death were increased after recovery following 60-min OGD. The increases were attenuated by pretreatment with antisense oligonucleotides against PSD-95 before OGD, but not by missense oligonucleotides or vehicle. PSD-95 antisense oligonucleotides also inhibited the increased interaction between PSD-95 and NR2A or Src, while NR2A expression did not change under this condition.

Conclusion

PSD-95 may be involved in regulating NR2A tyrosine phosphorylation by Src kinase. Inhibition of PSD-95 expression can be neuroprotective against apoptosislike cell death after recovery from OGD.

Keywords: postsynaptic density protein 95, N-methyl-D-aspartate receptor, oxygen-glucose deprivation, tyrosine phosphorylation, Src, cortical neurons

Footnotes

These authors contributed equally to this work.

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