High Glucose-Induced TRPC6 Channel Activation Decreases Glutamate Uptake in Rat Retinal Müller Cells

High glucose (HG) increases the production of reactive oxygen species (ROS), leading to decreased glutamate uptake in Müller cells. The transient receptor potential cation channel 6 (TRPC6) channel, an oxidative stress-sensitive Ca2+-permeable cationic channel, is readily detected in Müller cells an...

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Main Authors: Mingming Ma, Shuzhi Zhao, Jian Zhang, Tao Sun, Ying Fan, Zhi Zheng
Format: Article
Language:English
Published: Frontiers Media S.A. 2020-02-01
Series:Frontiers in Pharmacology
Subjects:
Online Access:https://www.frontiersin.org/article/10.3389/fphar.2019.01668/full
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language English
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author Mingming Ma
Mingming Ma
Mingming Ma
Mingming Ma
Mingming Ma
Shuzhi Zhao
Shuzhi Zhao
Shuzhi Zhao
Shuzhi Zhao
Shuzhi Zhao
Jian Zhang
Jian Zhang
Jian Zhang
Jian Zhang
Jian Zhang
Tao Sun
Tao Sun
Tao Sun
Tao Sun
Tao Sun
Ying Fan
Ying Fan
Ying Fan
Ying Fan
Ying Fan
Zhi Zheng
Zhi Zheng
Zhi Zheng
Zhi Zheng
Zhi Zheng
spellingShingle Mingming Ma
Mingming Ma
Mingming Ma
Mingming Ma
Mingming Ma
Shuzhi Zhao
Shuzhi Zhao
Shuzhi Zhao
Shuzhi Zhao
Shuzhi Zhao
Jian Zhang
Jian Zhang
Jian Zhang
Jian Zhang
Jian Zhang
Tao Sun
Tao Sun
Tao Sun
Tao Sun
Tao Sun
Ying Fan
Ying Fan
Ying Fan
Ying Fan
Ying Fan
Zhi Zheng
Zhi Zheng
Zhi Zheng
Zhi Zheng
Zhi Zheng
High Glucose-Induced TRPC6 Channel Activation Decreases Glutamate Uptake in Rat Retinal Müller Cells
Frontiers in Pharmacology
TRPC6 channel
diabetic retinopathy
Müller cells
glutamate uptake
reactive oxygen species
author_facet Mingming Ma
Mingming Ma
Mingming Ma
Mingming Ma
Mingming Ma
Shuzhi Zhao
Shuzhi Zhao
Shuzhi Zhao
Shuzhi Zhao
Shuzhi Zhao
Jian Zhang
Jian Zhang
Jian Zhang
Jian Zhang
Jian Zhang
Tao Sun
Tao Sun
Tao Sun
Tao Sun
Tao Sun
Ying Fan
Ying Fan
Ying Fan
Ying Fan
Ying Fan
Zhi Zheng
Zhi Zheng
Zhi Zheng
Zhi Zheng
Zhi Zheng
author_sort Mingming Ma
title High Glucose-Induced TRPC6 Channel Activation Decreases Glutamate Uptake in Rat Retinal Müller Cells
title_short High Glucose-Induced TRPC6 Channel Activation Decreases Glutamate Uptake in Rat Retinal Müller Cells
title_full High Glucose-Induced TRPC6 Channel Activation Decreases Glutamate Uptake in Rat Retinal Müller Cells
title_fullStr High Glucose-Induced TRPC6 Channel Activation Decreases Glutamate Uptake in Rat Retinal Müller Cells
title_full_unstemmed High Glucose-Induced TRPC6 Channel Activation Decreases Glutamate Uptake in Rat Retinal Müller Cells
title_sort high glucose-induced trpc6 channel activation decreases glutamate uptake in rat retinal müller cells
publisher Frontiers Media S.A.
series Frontiers in Pharmacology
issn 1663-9812
publishDate 2020-02-01
description High glucose (HG) increases the production of reactive oxygen species (ROS), leading to decreased glutamate uptake in Müller cells. The transient receptor potential cation channel 6 (TRPC6) channel, an oxidative stress-sensitive Ca2+-permeable cationic channel, is readily detected in Müller cells and highly expressed under HG conditions. Yet, the effect of high glucose-induced TRPC6 channel activation in Müller cells is poorly understood. We hypothesized that TRPC6 channel activation mediates high glucose-induced decreases in Müller cell glutamate uptake. We found RNA interference (RNAi) of the TRPC6 channel abolished HG-induced decreases in glutamate uptake and cell death. HG also decreased the expression of the glutamate-aspartate transporter (GLAST), which is the most important transporter involved in glutamate uptake. The mRNA level of ciliary neurotrophic factor (CNTF) in rMC-1 cells and the release of CNTF in the culture media was decreased, but the mRNA levels of IL-6 and vascular endothelial growth factor (VEGF) were increased under HG conditions. After RNAi silencing in rMC-1 cells, the mRNA levels of CNTF increased, but IL-6 and VEGF levels decreased. Furthermore, TRPC6 knockdown (KD) decreased expression of glial fibrillary acidic protein (GFAP) and increased expression of Kir4.1, pointing to inhibition of HG-induced gliosis in rMC-1 cells. ROS and intracellular Ca2+ levels decreased after TRPC6 knockdown. Exposure to Hyp9 (10 μM), a highly selective TRPC6 channel agonist, can aggravate HG-induced pathological changes. Collectively, our results suggest TRPC6 channel activation is involved in HG-induced decreases in glutamate uptake in rMC-1 cells. These findings provide novel insights into the role of TRPC6 in HG-induced retinal neurovasculopathy and suggest TRPC6 is a promising target for drug development for diabetic retinopathy (DR).
topic TRPC6 channel
diabetic retinopathy
Müller cells
glutamate uptake
reactive oxygen species
url https://www.frontiersin.org/article/10.3389/fphar.2019.01668/full
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spelling doaj-9ac77e3f22d34dafa3b4bc4ba1333e872020-11-25T02:50:24ZengFrontiers Media S.A.Frontiers in Pharmacology1663-98122020-02-011010.3389/fphar.2019.01668498451High Glucose-Induced TRPC6 Channel Activation Decreases Glutamate Uptake in Rat Retinal Müller CellsMingming Ma0Mingming Ma1Mingming Ma2Mingming Ma3Mingming Ma4Shuzhi Zhao5Shuzhi Zhao6Shuzhi Zhao7Shuzhi Zhao8Shuzhi Zhao9Jian Zhang10Jian Zhang11Jian Zhang12Jian Zhang13Jian Zhang14Tao Sun15Tao Sun16Tao Sun17Tao Sun18Tao Sun19Ying Fan20Ying Fan21Ying Fan22Ying Fan23Ying Fan24Zhi Zheng25Zhi Zheng26Zhi Zheng27Zhi Zheng28Zhi Zheng29Department of Ophthalmology, Shanghai General Hospital, Shanghai, ChinaNational Clinical Research Center for Eye Diseases, Shanghai, ChinaShanghai Key Laboratory of Ocular Fundus Diseases, Shanghai Jiao Tong University, Shanghai, ChinaShanghai Engineering Center for Visual Science and Photomedicine, Shanghai General Hospital, Shanghai Jiao Tong University, Shanghai, ChinaShanghai Engineering Center for Precise Diagnosis and Treatment of Eye Diseases, Shanghai, ChinaDepartment of Ophthalmology, Shanghai General Hospital, Shanghai, ChinaNational Clinical Research Center for Eye Diseases, Shanghai, ChinaShanghai Key Laboratory of Ocular Fundus Diseases, Shanghai Jiao Tong University, Shanghai, ChinaShanghai Engineering Center for Visual Science and Photomedicine, Shanghai General Hospital, Shanghai Jiao Tong University, Shanghai, ChinaShanghai Engineering Center for Precise Diagnosis and Treatment of Eye Diseases, Shanghai, ChinaDepartment of Ophthalmology, Shanghai General Hospital, Shanghai, ChinaNational Clinical Research Center for Eye Diseases, Shanghai, ChinaShanghai Key Laboratory of Ocular Fundus Diseases, Shanghai Jiao Tong University, Shanghai, ChinaShanghai Engineering Center for Visual Science and Photomedicine, Shanghai General Hospital, Shanghai Jiao Tong University, Shanghai, ChinaShanghai Engineering Center for Precise Diagnosis and Treatment of Eye Diseases, Shanghai, ChinaDepartment of Ophthalmology, Shanghai General Hospital, Shanghai, ChinaNational Clinical Research Center for Eye Diseases, Shanghai, ChinaShanghai Key Laboratory of Ocular Fundus Diseases, Shanghai Jiao Tong University, Shanghai, ChinaShanghai Engineering Center for Visual Science and Photomedicine, Shanghai General Hospital, Shanghai Jiao Tong University, Shanghai, ChinaShanghai Engineering Center for Precise Diagnosis and Treatment of Eye Diseases, Shanghai, ChinaDepartment of Ophthalmology, Shanghai General Hospital, Shanghai, ChinaNational Clinical Research Center for Eye Diseases, Shanghai, ChinaShanghai Key Laboratory of Ocular Fundus Diseases, Shanghai Jiao Tong University, Shanghai, ChinaShanghai Engineering Center for Visual Science and Photomedicine, Shanghai General Hospital, Shanghai Jiao Tong University, Shanghai, ChinaShanghai Engineering Center for Precise Diagnosis and Treatment of Eye Diseases, Shanghai, ChinaDepartment of Ophthalmology, Shanghai General Hospital, Shanghai, ChinaNational Clinical Research Center for Eye Diseases, Shanghai, ChinaShanghai Key Laboratory of Ocular Fundus Diseases, Shanghai Jiao Tong University, Shanghai, ChinaShanghai Engineering Center for Visual Science and Photomedicine, Shanghai General Hospital, Shanghai Jiao Tong University, Shanghai, ChinaShanghai Engineering Center for Precise Diagnosis and Treatment of Eye Diseases, Shanghai, ChinaHigh glucose (HG) increases the production of reactive oxygen species (ROS), leading to decreased glutamate uptake in Müller cells. The transient receptor potential cation channel 6 (TRPC6) channel, an oxidative stress-sensitive Ca2+-permeable cationic channel, is readily detected in Müller cells and highly expressed under HG conditions. Yet, the effect of high glucose-induced TRPC6 channel activation in Müller cells is poorly understood. We hypothesized that TRPC6 channel activation mediates high glucose-induced decreases in Müller cell glutamate uptake. We found RNA interference (RNAi) of the TRPC6 channel abolished HG-induced decreases in glutamate uptake and cell death. HG also decreased the expression of the glutamate-aspartate transporter (GLAST), which is the most important transporter involved in glutamate uptake. The mRNA level of ciliary neurotrophic factor (CNTF) in rMC-1 cells and the release of CNTF in the culture media was decreased, but the mRNA levels of IL-6 and vascular endothelial growth factor (VEGF) were increased under HG conditions. After RNAi silencing in rMC-1 cells, the mRNA levels of CNTF increased, but IL-6 and VEGF levels decreased. Furthermore, TRPC6 knockdown (KD) decreased expression of glial fibrillary acidic protein (GFAP) and increased expression of Kir4.1, pointing to inhibition of HG-induced gliosis in rMC-1 cells. ROS and intracellular Ca2+ levels decreased after TRPC6 knockdown. Exposure to Hyp9 (10 μM), a highly selective TRPC6 channel agonist, can aggravate HG-induced pathological changes. Collectively, our results suggest TRPC6 channel activation is involved in HG-induced decreases in glutamate uptake in rMC-1 cells. These findings provide novel insights into the role of TRPC6 in HG-induced retinal neurovasculopathy and suggest TRPC6 is a promising target for drug development for diabetic retinopathy (DR).https://www.frontiersin.org/article/10.3389/fphar.2019.01668/fullTRPC6 channeldiabetic retinopathyMüller cellsglutamate uptakereactive oxygen species