Effects of activating mTORC2/Akt signaling pathway on dopaminergic neurons and behaviors in 6-hydroxydopamin model mice

BIAN Wei; LI Meng-Yi; ZHOU Peng; LI Jun-Wei; ZHANG Ting; WU An-Ting; QI Shuang-Shuang; CUI Huai-Rui; SUN Chen-You

doi:10.16098/j.issn.0529-1356.2023.01.002

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Acta Anatomica Sinica ›› 2023, Vol. 54 ›› Issue (1) : 13-22. DOI: 10.16098/j.issn.0529-1356.2023.01.002

Neurobiology

Effects of activating mTORC2/Akt signaling pathway on dopaminergic neurons and behaviors in 6-hydroxydopamin model mice

BIAN Wei^1,2 LI Meng-yi^1,2 ZHOU Peng^1,2 LI Jun-wei^1,2 ZHANG Ting³ WU An-ting^1,2 QI Shuang-shuang⁴ CUI Huai-rui¹ SUN Chen-you^1,2*

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Abstract

Objective To explore the effect of activation of mammalian target of rapmycin complex 2(mTORC2)/Akt signaling pathway on dopaminergic neurons and behavior in 6-hydroxydopamine (6-OHDA) model mice and its possible mechanism. Methods Selecting 36 mice which The Nestin-CreERTM and ROSA26-LacZ reporter genes were detected at the same time in 3-month-old male C57BL/6J mice weighing 20-25 g divideng them into 4 gruops, NS+ corn oil group, 6-OHDA+corn oil group, 6-OHDA+PP242 group and 6-OHDA+A-443654 group, and 6-OHDA was injected into the right striatum of the brain to replicate the Parkinson’s disease (PD) model of mice, and then daily intraperitoneal injection of mTORC2/Akt signaling pathway agonist A-443654 or inhibitor PP242. Serum interleukin-1β (IL-1β) and tumor necrosis factor-α(TNF-α)levels were measured by enzymelinked immunosorbent assay. Immunohistochemistry and immunofluorescence staining were performed to investigate the change of microglia, dopaminergic neurons as well as neural progenitor cells (NPCs). Western blotting was used to detect the expression of related protein of mTORC2/Akt signaling pathway including rictor, p-Akt and regulated in development and DNA dgmage responses 1(REDD1) and the interaction between them were verified by immunoprecipitation. Finally, the behavioral performance of each group of mice was observed. Results With the activation of microglia and the increase of inflammatory factors in PD model mice, the number of dopaminergic neurons in the substantia nigra(SN) decreased significantly, and the motor function of the mice was impaired, but the number of NPCs increased significantly compared with the control mice, mTORC2/Akt signaling pathway related protein expression was also significantly up-regulated. A-443654 treatment further up-regulated the expression of these proteins, meanwhile the indicators mentioned above were ameliorated. However, the inhibitor PP242 treatment group showed completely opposite result with the agonist group. Conclusion A-443654 can promote the proliferation of NPCs and the number of new-born dopaminergic neurons by up-regulating related proteins of mTORC2/Akt signaling pathway, and reducing the activation of microglia and the level of inflammation factors, which ultimately lead to the amelioration of SN-striatal dopaminergic neurons and behavioral performance in PD model mice.

Key words

Mammalian target of rapamycin complex 2

/ Parkinson’s disease / Dopaminergic neuron / Microglial cell / Nerve regeneration / Western blotting / Mouse

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BIAN Wei LI Meng-yi ZHOU Peng LI Jun-wei ZHANG Ting WU An-ting QI Shuang-shuang CUI Huai-rui SUN Chen-you. Effects of activating mTORC2/Akt signaling pathway on dopaminergic neurons and behaviors in 6-hydroxydopamin model mice[J]. Acta Anatomica Sinica. 2023, 54(1): 13-22 https://doi.org/10.16098/j.issn.0529-1356.2023.01.002

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