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Overexpression of G Protein-Coupled Receptor Kinase 2 Inhibits Neuronal Electrophysiological Activity in SH-SY5Y Cells via Blocking Na Channels; A Laboratory Study
Vol 36, Issue 4, 2022
Abstract
Background: G protein-coupled receptor kinase 2 (GRK2) is involved in G protein-coupled receptor (GPCRs) pathway and non-receptor signaling pathway, which is only the non-receptor signaling pathway or both the receptor and non-receptor signaling pathway closely related to the occurrence of neurological diseases. However, the regulatory mechanism of GRK2 in electrophysiological activity is still unclear. Methods: In this study, GRK2 shRNA plasmid or GRK2 overexpression plasmid was transfected into human neuroblastoma cells SH-SY5Y. Transfection efficiency was detected using western blot and patch-clamp techniques were used to measure changes in electrophysiological activity in SH-SY5Y cells after overexpression or knockdown of GRK2. Results: Western blot results showed successful transfection. Additionally, the measurement by patch-clamp technique demonstrated that GRK2 overexpression significantly decreased both action potential firing frequency and Na+ current, which inhibited neuronal excitability in SH-SY5Y cells (p < 0.05). However, knockdown of GRK2 had no significant effect on action potential firing frequency and Na+ current in SH-SY5Y cells (p > 0.05). Conclusions: GRK2 overexpression may suppress the electrophysiological activity of SH-SY5Y cells by blocking the Na channel.
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Copyright (c) 2022 Yuan-yuan Ma, Ye Xu, Tian Zhang, Zhi-bin Chen, Tan Wang
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Medical Genetics, University of Torino Medical School, Italy

Department of Biomedical, Surgical and Dental Sciences, University of Milan, Italy