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Blocking mPTP on Neural Stem Cells and Activating the Nicotinic Acetylcholine Receptor α7 Subunit on Microglia Attenuate Aβ-Induced Neurotoxicity on Neural Stem Cells

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Abstract

β-Amyloid (Aβ) can stimulate microglia to release a variety of proinflammatory cytokines and induce neurotoxicity. Nicotine has been reported to inhibit TNF-α, IL-1, and ROS production in microglia. Mitochondrial permeability transition pore (mPTP) plays an important role in neurotoxicity as well. Here, we investigated whether activating the microglial α7-nAChR has a neuroprotective role on neural stem cells (NSCs) and the function of mPTP in NSCs in this process. The expression of α7-nAChR in rat NSCs was detected by immunocytochemistry and RT-PCR. The viability of microglia and NSCs was examined by MTT assay. The mitochondrial membrane potential (ΔΨm) and morphological characteristics of NSCs was measured by JC-1 staining and transmission electron microscopy respectively. The distribution of cytochrome c in the subcellular regions of NSCs was visualized by confocal laser scanning microscopy, and the expression levels of cyclophilin D and cleaved caspase-3 were assayed by western blot. The apoptotic rate of NSCs was measured by flow cytometry. The expression of α7-nAChR was detected in microglial cells, but no expression was found in NSCs. The viability of rat microglial cells and NSCs was not affected by reagents or coculture itself. Aβ1–42-mediated microglial activation impaired the morphology and the ΔΨm of mitochondria of NSCs as well as increased cell apoptosis. However, the damage was attenuated when the α7-nAChRs on microglial cells were activated or the mPTPs on NSCs were blocked. Blockade of mPTPs on NSCs and activation of α7-nAChRs on microglia exhibit neuroprotective roles in Aβ-induced neurotoxicity of NSCs.

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Abbreviations

AD:

Alzheimer’s disease

Aβ:

β-Amyloid

CsA:

Cyclosporine A

CypD:

Cyclophilin D

IL-1:

Interleukin-1

ΔΨm:

Mitochondrial membrane potential

mPTP:

Mitochondrial permeability transition pore

NSCs:

Neural stem cells

α7-nAChR:

Nicotinic acetylcholine receptor α7 subunit

TNF-α:

Tumor necrosis factor alpha

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Acknowledgments

This work was supported in part by grants from the National Natural Science Foundation of China (No. 30973162) and the National Natural Science Foundation of Guangdong Province (No. S2013010015546). In addition, this work was supported in part by the Guangdong Provincial Key Laboratory of Malignant Tumor Epigenetics and Gene Regulation, Sun Yat-Sen Memorial Hospital, Sun Yat-Sen University.

Authors’ Contributions

YW§, QC, and SJ designed the research. QC and DJ conducted the analysis. QC, DJ, KW, ML, YW, and NZ contributed to the MTT assay, western blotting, and immunofluorescence experiments. QC, XX, and YW performed the RT-PCR experiments. QC and YW performed the flow cytometry and cell culture experiments. QC performed the electron microscopy analyses. YW§, QC, and SJ analyzed the data and wrote the paper.

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    Authors and Affiliations

    1. Department of Pharmacy, Southern Medical University, Zhujiang Hospital, Guangzhou, China

      Qingzhuang Chen, Deqi Jiang, Yan Wang, Xiaodan Xiao, Ning Zhu, Mingxing Li, Siyuan Jia & Yong Wang

    2. Department of Clinical Pharmacy, Guangzhou Hospital of Integrated Traditional and Western Medicine, Guangzhou, China

      Qingzhuang Chen

    3. Department of Neurosurgery, Nanfang Hospital, Southern Medical University, Guangzhou, China

      Kewan Wang

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    Qingzhuang Chen and Kewan Wang have contributed equally to this work.

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    Chen, Q., Wang, K., Jiang, D. et al. Blocking mPTP on Neural Stem Cells and Activating the Nicotinic Acetylcholine Receptor α7 Subunit on Microglia Attenuate Aβ-Induced Neurotoxicity on Neural Stem Cells. Neurochem Res 41, 1483–1495 (2016). https://doi.org/10.1007/s11064-016-1862-8

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