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

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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.

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Authors and Affiliations

  1. Jiangsu Key Laboratory of Brain Disease Bioinformation, Xuzhou Medical College, Xuzhou, 221002, China

    Jing-Zhi Yan, Yong Liu, Yan-Yan Zong & Guang-Yi Zhang

  2. Research Center for Biochemistry and Molecular Biology, Xuzhou Medical College, Xuzhou, 221002, China

    Jing-Zhi Yan, Yong Liu, Yan-Yan Zong & Guang-Yi Zhang

Authors
  1. Jing-Zhi Yan
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  2. Yong Liu
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  3. Yan-Yan Zong
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  4. Guang-Yi Zhang
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Correspondence to Guang-Yi Zhang.

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These authors contributed equally to this work.

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Yan, JZ., Liu, Y., Zong, YY. et al. Knock-down of postsynaptic density protein 95 expression by antisense oligonucleotides protects against apoptosis-like cell death induced by oxygen-glucose deprivation in vitro . Neurosci. Bull. 28, 69–76 (2012). https://doi.org/10.1007/s12264-012-1065-5

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  • DOI: https://doi.org/10.1007/s12264-012-1065-5

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