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...
Main Authors: | , , , , , |
---|---|
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 |
id |
doaj-9ac77e3f22d34dafa3b4bc4ba1333e87 |
---|---|
record_format |
Article |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
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 |
work_keys_str_mv |
AT mingmingma highglucoseinducedtrpc6channelactivationdecreasesglutamateuptakeinratretinalmullercells AT mingmingma highglucoseinducedtrpc6channelactivationdecreasesglutamateuptakeinratretinalmullercells AT mingmingma highglucoseinducedtrpc6channelactivationdecreasesglutamateuptakeinratretinalmullercells AT mingmingma highglucoseinducedtrpc6channelactivationdecreasesglutamateuptakeinratretinalmullercells AT mingmingma highglucoseinducedtrpc6channelactivationdecreasesglutamateuptakeinratretinalmullercells AT shuzhizhao highglucoseinducedtrpc6channelactivationdecreasesglutamateuptakeinratretinalmullercells AT shuzhizhao highglucoseinducedtrpc6channelactivationdecreasesglutamateuptakeinratretinalmullercells AT shuzhizhao highglucoseinducedtrpc6channelactivationdecreasesglutamateuptakeinratretinalmullercells AT shuzhizhao highglucoseinducedtrpc6channelactivationdecreasesglutamateuptakeinratretinalmullercells AT shuzhizhao highglucoseinducedtrpc6channelactivationdecreasesglutamateuptakeinratretinalmullercells AT jianzhang highglucoseinducedtrpc6channelactivationdecreasesglutamateuptakeinratretinalmullercells AT jianzhang highglucoseinducedtrpc6channelactivationdecreasesglutamateuptakeinratretinalmullercells AT jianzhang highglucoseinducedtrpc6channelactivationdecreasesglutamateuptakeinratretinalmullercells AT jianzhang highglucoseinducedtrpc6channelactivationdecreasesglutamateuptakeinratretinalmullercells AT jianzhang highglucoseinducedtrpc6channelactivationdecreasesglutamateuptakeinratretinalmullercells AT taosun highglucoseinducedtrpc6channelactivationdecreasesglutamateuptakeinratretinalmullercells AT taosun highglucoseinducedtrpc6channelactivationdecreasesglutamateuptakeinratretinalmullercells AT taosun highglucoseinducedtrpc6channelactivationdecreasesglutamateuptakeinratretinalmullercells AT taosun highglucoseinducedtrpc6channelactivationdecreasesglutamateuptakeinratretinalmullercells AT taosun highglucoseinducedtrpc6channelactivationdecreasesglutamateuptakeinratretinalmullercells AT yingfan highglucoseinducedtrpc6channelactivationdecreasesglutamateuptakeinratretinalmullercells AT yingfan highglucoseinducedtrpc6channelactivationdecreasesglutamateuptakeinratretinalmullercells AT yingfan highglucoseinducedtrpc6channelactivationdecreasesglutamateuptakeinratretinalmullercells AT yingfan highglucoseinducedtrpc6channelactivationdecreasesglutamateuptakeinratretinalmullercells AT yingfan highglucoseinducedtrpc6channelactivationdecreasesglutamateuptakeinratretinalmullercells AT zhizheng highglucoseinducedtrpc6channelactivationdecreasesglutamateuptakeinratretinalmullercells AT zhizheng highglucoseinducedtrpc6channelactivationdecreasesglutamateuptakeinratretinalmullercells AT zhizheng highglucoseinducedtrpc6channelactivationdecreasesglutamateuptakeinratretinalmullercells AT zhizheng highglucoseinducedtrpc6channelactivationdecreasesglutamateuptakeinratretinalmullercells AT zhizheng highglucoseinducedtrpc6channelactivationdecreasesglutamateuptakeinratretinalmullercells |
_version_ |
1724738845890576384 |
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 |