The decrease of GluN2B and its phosphorylation at Tyr-1336 in extrasynaptic subunits is associated with neuroprotection induced by hypoxia preconditioning

Prior research has firmly established that the N-methyl-d-aspartate (NMDA) receptor subunit 2 B (GluN2B) and its phosphorylation contribute to ischemic/hypoxic brain injury. Hypoxic preconditioning (HPC) is an endogenous mechanism that protects the brain from both ischaemic and hypoxic damage. In th...

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Main Authors: Haidong Hou, Jing Yang, Gang Fu, Xiaolei Liu, You Liu, Kerui Gong, Ruifang Qi, Xiangning Jiang, Guo Shao
Format: Article
Language:English
Published: Elsevier 2025-07-01
Series:Brain Research Bulletin
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Online Access:http://www.sciencedirect.com/science/article/pii/S0361923025002126
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author Haidong Hou
Jing Yang
Gang Fu
Xiaolei Liu
You Liu
Kerui Gong
Ruifang Qi
Xiangning Jiang
Guo Shao
author_facet Haidong Hou
Jing Yang
Gang Fu
Xiaolei Liu
You Liu
Kerui Gong
Ruifang Qi
Xiangning Jiang
Guo Shao
author_sort Haidong Hou
collection DOAJ
description Prior research has firmly established that the N-methyl-d-aspartate (NMDA) receptor subunit 2 B (GluN2B) and its phosphorylation contribute to ischemic/hypoxic brain injury. Hypoxic preconditioning (HPC) is an endogenous mechanism that protects the brain from both ischaemic and hypoxic damage. In this study, we explored the effects of HPC on GluN2B and its phosphorylation at two sites (tyrosine residues 1252 and 1336), catalysed by Fyn, in the hippocampus both in vivo and in vitro. Animal and cellular models of HPC were developed by subjecting mice and the mouse hippocampal neuronal cell line HT22 to repeated hypoxia. Levels of GluN2B and its phosphorylation at the tyrosine residues 1336 (pY1336 GluN2B) and 1252 (pY1252 GluN2B) were detected in HPC-treated hippocampi and HT22 cells using western blotting and immunofluorescence. The distributions of GluN2B, pY1336 GluN2B, and pY1252 GluN2B in the synaptic (TxP) and extrasynaptic components (TxS) were analysed by western blotting. Caspase-3 and spectrin, both markers of cellular injury, were further measured using western blotting. HPC downregulated GluN2B and pY1336 GluN2B levels in the hippocampus and HT22 cells. The changes in GluN2B and pY1336 GluN2B levels in the extrasynaptic components were similar to those in the hippocampus and HT22 cells, while the changes in the synaptic components showed the opposite trend which increased after HPC. The downregulation of GluN2B and pY1336 GluN2B may be associated with neuroprotection induced by HPC. Additionally, their localization at synaptic and extrasynaptic sites may play distinct roles in neuroprotection.
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spelling doaj-art-c9a54e44287241da8e1dc3e8bfaceb3b2025-08-20T03:46:34ZengElsevierBrain Research Bulletin1873-27472025-07-0122711140010.1016/j.brainresbull.2025.111400The decrease of GluN2B and its phosphorylation at Tyr-1336 in extrasynaptic subunits is associated with neuroprotection induced by hypoxia preconditioningHaidong Hou0Jing Yang1Gang Fu2Xiaolei Liu3You Liu4Kerui Gong5Ruifang Qi6Xiangning Jiang7Guo Shao8Department of Neurosurgery, Dongguan Qingxi Hospital, Dongguan, PR ChinaInner Mongolia Key Laboratory of Hypoxic Translational Medicine, Baotou Medical College, Baotou, PR China; Beijing Key Laboratory of Hypoxic Conditioning Translational Medicine, Xuanwu Hospital, Capital Medical University, Beijing, PR ChinaLonggang Institute of Medical Imaging, Shantou University Medical College & The Third People's Hospital of Longgang District, Shenzhen, PR ChinaInner Mongolia Key Laboratory of Hypoxic Translational Medicine, Baotou Medical College, Baotou, PR China; Beijing Key Laboratory of Hypoxic Conditioning Translational Medicine, Xuanwu Hospital, Capital Medical University, Beijing, PR ChinaInner Mongolia Key Laboratory of Hypoxic Translational Medicine, Baotou Medical College, Baotou, PR China; Beijing Key Laboratory of Hypoxic Conditioning Translational Medicine, Xuanwu Hospital, Capital Medical University, Beijing, PR ChinaDepartment of Oral and Maxillofacial Surgery, University of California San Francisco, San Francisco, USAInner Mongolia Key Laboratory of Hypoxic Translational Medicine, Baotou Medical College, Baotou, PR China; Beijing Key Laboratory of Hypoxic Conditioning Translational Medicine, Xuanwu Hospital, Capital Medical University, Beijing, PR China; Corresponding authors at: Inner Mongolia Key Laboratory of Hypoxic Translational Medicine, Baotou Medical College, Baotou, PR China.Department of Neurology, University of California San Francisco, San Francisco, CA, USA; Corresponding author.Inner Mongolia Key Laboratory of Hypoxic Translational Medicine, Baotou Medical College, Baotou, PR China; Beijing Key Laboratory of Hypoxic Conditioning Translational Medicine, Xuanwu Hospital, Capital Medical University, Beijing, PR China; Longgang Institute of Medical Imaging, Shantou University Medical College & The Third People's Hospital of Longgang District, Shenzhen, PR China; Corresponding authors at: Inner Mongolia Key Laboratory of Hypoxic Translational Medicine, Baotou Medical College, Baotou, PR China.Prior research has firmly established that the N-methyl-d-aspartate (NMDA) receptor subunit 2 B (GluN2B) and its phosphorylation contribute to ischemic/hypoxic brain injury. Hypoxic preconditioning (HPC) is an endogenous mechanism that protects the brain from both ischaemic and hypoxic damage. In this study, we explored the effects of HPC on GluN2B and its phosphorylation at two sites (tyrosine residues 1252 and 1336), catalysed by Fyn, in the hippocampus both in vivo and in vitro. Animal and cellular models of HPC were developed by subjecting mice and the mouse hippocampal neuronal cell line HT22 to repeated hypoxia. Levels of GluN2B and its phosphorylation at the tyrosine residues 1336 (pY1336 GluN2B) and 1252 (pY1252 GluN2B) were detected in HPC-treated hippocampi and HT22 cells using western blotting and immunofluorescence. The distributions of GluN2B, pY1336 GluN2B, and pY1252 GluN2B in the synaptic (TxP) and extrasynaptic components (TxS) were analysed by western blotting. Caspase-3 and spectrin, both markers of cellular injury, were further measured using western blotting. HPC downregulated GluN2B and pY1336 GluN2B levels in the hippocampus and HT22 cells. The changes in GluN2B and pY1336 GluN2B levels in the extrasynaptic components were similar to those in the hippocampus and HT22 cells, while the changes in the synaptic components showed the opposite trend which increased after HPC. The downregulation of GluN2B and pY1336 GluN2B may be associated with neuroprotection induced by HPC. Additionally, their localization at synaptic and extrasynaptic sites may play distinct roles in neuroprotection.http://www.sciencedirect.com/science/article/pii/S0361923025002126Hypoxic preconditioningNeuroprotectionGluN2BSynapse
spellingShingle Haidong Hou
Jing Yang
Gang Fu
Xiaolei Liu
You Liu
Kerui Gong
Ruifang Qi
Xiangning Jiang
Guo Shao
The decrease of GluN2B and its phosphorylation at Tyr-1336 in extrasynaptic subunits is associated with neuroprotection induced by hypoxia preconditioning
Brain Research Bulletin
Hypoxic preconditioning
Neuroprotection
GluN2B
Synapse
title The decrease of GluN2B and its phosphorylation at Tyr-1336 in extrasynaptic subunits is associated with neuroprotection induced by hypoxia preconditioning
title_full The decrease of GluN2B and its phosphorylation at Tyr-1336 in extrasynaptic subunits is associated with neuroprotection induced by hypoxia preconditioning
title_fullStr The decrease of GluN2B and its phosphorylation at Tyr-1336 in extrasynaptic subunits is associated with neuroprotection induced by hypoxia preconditioning
title_full_unstemmed The decrease of GluN2B and its phosphorylation at Tyr-1336 in extrasynaptic subunits is associated with neuroprotection induced by hypoxia preconditioning
title_short The decrease of GluN2B and its phosphorylation at Tyr-1336 in extrasynaptic subunits is associated with neuroprotection induced by hypoxia preconditioning
title_sort decrease of glun2b and its phosphorylation at tyr 1336 in extrasynaptic subunits is associated with neuroprotection induced by hypoxia preconditioning
topic Hypoxic preconditioning
Neuroprotection
GluN2B
Synapse
url http://www.sciencedirect.com/science/article/pii/S0361923025002126
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