Abstract
Increasing evidence has indicated that the Golgi apparatus (GA) is involved in the development of cerebral ischemia–reperfusion (IR) injury. Finding effective neuroprotective agents targeting GA has become a priority in the treatment of ischemic stroke. GM130, a key structural protein present on the cis-face of the GA, maintains its structure through its phosphorylation and dephosphorylation. However, the molecular mechanisms by which GM130 regulates IR-induced neuronal apoptosis are not well elucidated. Mouse neuroblastoma Neuro2a (N2A) cells were subjected to oxygen–glucose deprivation and reperfusion (OGDR) insult. Cell proliferation and apoptosis were determined using MTT assay, TUNEL staining, and flow cytometry. GA morphology was detected by immunocytochemical staining and immunofluorescence microscopy. GA-related protein and mRNA levels were detected by WB and qPCR, respectively. Treatment with Purvalanol A, an effective Cdk1 inhibitor, and transfection of Cdk1-shRNA were carried out to inhibit OGDR-induced Cdk1 elevation. The results demonstrated that OGDR induced Golgi fragmentation, neuronal apoptosis, GM130 phosphorylation, and p115 cleavage in N2A cells. Cdk1 elevation after OGDR was closely correlated with GM130 phosphorylation, not p115. Inhibition of Cdk1 significantly attenuated OGDR-induced Golgi fragmentation and cell apoptosis. Cdk1 interacted with GM130 and decreased its phosphorylation on the serine 25 site in N2A cells exposed to OGDR. The present findings reveal that Cdk1 protects against IR-induced GA fragmentation and apoptosis, likely through the mediation of GM130 phosphorylation. This neuroprotective potential of Cdk1 against IR insult and the underlying mechanism will pave the way for potential clinical applications targeting the GA organelle for cerebral IR-related disorders.
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This work was supported by the National Natural Science Foundation of China (Grant No. 82002010).
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All authors contributed to the study conception and design. Material preparation, implementation of the experiment, data collection and analysis were performed by ZL, WZ and JX. The first draft of the manuscript was written by XM and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.
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Li, Z., Zhang, W., Xu, J. et al. Cdk1 protects against oxygen-glucose deprivation and reperfusion-induced Golgi fragmentation and apoptosis through mediating GM130 phosphorylation. J Mol Histol 54, 609–619 (2023). https://doi.org/10.1007/s10735-023-10164-3
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DOI: https://doi.org/10.1007/s10735-023-10164-3