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MCT4-Mediated Expression of EAAT1 is Involved in the Resistance to Hypoxia Injury in Astrocyte–Neuron co-Cultures

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Abstract

Hypoxic stressors contribute to neuronal death in many brain diseases. Astrocyte processes surround most neurons and are therefore anatomically well-positioned to shield them from hypoxic injury. Excitatory amino acid transporters (EAATs), represent the sole mechanism of active reuptake of glutamate into the astrocytes and neurons and are essential to dampen neuronal excitation following glutamate release at synapses. Glutamate clearance impairment from any factors is bound to result in an increase in hypoxic neuronal injury. The brain energy metabolism under hypoxic conditions depends on monocarboxylate transporters (MCTs) that are expressed by neurons and glia. Previous co-immunoprecipitation experiments revealed that MCT4 directly modulate EAAT1 in astrocytes. The reduction in both surface proteins may act synergistically to induce neuronal hyperexcitability and excitotoxicity. Therefore we hypothesized that astrocytes would respond to hypoxic conditions by enhancing their expression of MCT4 and EAAT1, which, in turn, would enable them to better support neurons to survive lethal hypoxia injury. An oxygen deprivation (OD) protocol was used in primary cultures of neurons, astrocytes, and astrocytes–neurons derived from rat hippocampus, with or without MCT4-targeted short hairpin RNA (shRNA) transfection. Cell survival, expression of MCT4, EAAT1, glial fibrillary acidic protein and neuronal nuclear antigen were evaluated. OD resulted in significant cell death in neuronal cultures and up-regulation of MCT4, EAAT1 expression respectively in primary cell cultures, but no injury in neuron–astrocyte co-cultures and astrocyte cultures. However, neuronal cell death in co-cultures was increased exposure to shRNA-MCT4 prior to OD. These findings demonstrate that the MCT4-mediated expression of EAAT1 is involved in the resistance to hypoxia injury in astrocyte–neuron co-cultures.

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Acknowledgments

The authors thank M.M. Qiang Zhang for help with the primary cell culture protocol and Yuhu Liu for his contributions to this work.

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    Authors and Affiliations

    1. Department of Neurosurgery, AnNing Branch Hospital, Lanzhou General Hospital of Lanzhou Military Command, Lanzhou, 730070, Gansu Province, China

      Chen Gao

    2. Department of Orthopedics, AnNing Branch Hospital, Lanzhou General Hospital of Lanzhou Military Command, Lanzhou, 730070, Gansu Province, China

      Wenxia Zhu

    3. Department of Neurosurgery, Lanzhou General Hospital of Lanzhou Military Command, Lanzhou, 730050, Gansu Province, China

      Lizhuang Tian & Zhiyun Li

    4. Laboratory Animal Center of Lanzhou University, Lanzhou, 730000, Gansu Province, China

      Jingke Zhang

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    Correspondence to Zhiyun Li.

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    Chen Gao and Wenxia Zhu Contributed equally to this work.

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    Gao, C., Zhu, W., Tian, L. et al. MCT4-Mediated Expression of EAAT1 is Involved in the Resistance to Hypoxia Injury in Astrocyte–Neuron co-Cultures. Neurochem Res 40, 818–828 (2015). https://doi.org/10.1007/s11064-015-1532-2

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