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Proteasome Inhibition-Induced Downregulation of Akt/GSK-3β Pathway Contributes to Abnormality of Tau in Hippocampal Slice

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Abstract

Proteasome inhibition can induce abnormal accumulation and phosphorylation of microtubule-associated protein tau. The major function of tau protein is to promote microtubules assembly and stabilization, and abnormal tau protein would disturb its microtubule-binding function. In this study, proteasome inhibitor MG132 was used to treat hippocampal slices to explore the role and mechanism of Akt/glycogen synthase kinase-3β (GSK-3β) in proteasome inhibition-induced tau abnormality. During the culture period, we measure the lactate dehydrogenase (LDH) content to assay the viability of hippocampal slices. Following 2.5 and 5 μM MG132 treatment for 6 h, we detected the expression, phosphorylation modification, and microtubule-binding function of tau protein of slices. We also analyzed the changed activities of glycogen synthase kinase-3β (GSK-3β) and protein kinase B (PKB/Akt) and the level of heat shock protein 90 (Hsp90) in the process. In addition, co-immunoprecipitation was used to investigate the interaction between Akt and Hsp90, Akt and protein phosphatase-2A (PP2A) in the MG132-treated organotypic hippocampal slices. Our results indicated that proteasome inhibition led to degradation obstacles and abnormal phosphorylation of tau protein. The downregulated Akt/GSK-3β signaling pathway might be responsible for the abnormal phosphorylation of tau protein at multiple sites which further reduced the microtubule-binding function of tau protein. Furthermore, proteasome inhibition decreased the binding capacity of Akt-Hsp90 while increased the Akt-PP2A binding ability which mediated Akt inactivity. This current study establishes a hippocampal slice model targeting Akt/GSK-3β signaling pathway to explore the pivotal role of proteasome inhibition in tau pathology.

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Acknowledgments

We thank Professor Wang Jianzhi at Tongji Medical College of Huazhong University of Sciences and Technology, Wuhan, China, for providing R134d used in this study. This work was supported by grants from the National Natural Science Foundation of China (No. 30700208, No. 30800329, and No. 31172102).

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The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. School of Life Sciences and Hubei Key Laboratory of Genetic Regulation and Integrative Biology, Central China Normal University, NO. 152, Luoyu Road, Wuhan, 430079, China

    Min Xie, Ruihong Shi, Ying Pan, Tao Zeng, Qicai Chen, Shaohui Wang & Xiaomei Liao

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  1. Min Xie
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Correspondence to Xiaomei Liao.

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Min Xie, Ruihong Shi and Ying Pan contributed equally to this work.

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Fig. S1

Effect of lactacystin induced proteasome inhibition on tau-Ser214 site phosphorylation in HEK293/tau441 cell line. The phosphorylation-dependent antibody pSer214 was used to measure the alteration of tau-Ser214 site, following lactacystin treatment for 24 h, the relative level of pS214 decreased in a dose-dependent manner. The results are expressed as the mean ± SD (n = 3); **p < 0.01, ***p < 0.001, lactacystin vs. controls. (GIF 45 kb)

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Xie, M., Shi, R., Pan, Y. et al. Proteasome Inhibition-Induced Downregulation of Akt/GSK-3β Pathway Contributes to Abnormality of Tau in Hippocampal Slice. Mol Neurobiol 50, 888–895 (2014). https://doi.org/10.1007/s12035-014-8702-0

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