High glucose induces mitochondrial dysfunction in retinal müller cells

Implications for diabetic retinopathy

Thomas Tien, Joyce Zhang, Tetsuya Muto, Dongjoon Kim, Vijay P Sarthy, Sayon Roy*

*Corresponding author for this work

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

PURPOSE. To investigate whether high glucose (HG) induces mitochondrial dysfunction and promotes apoptosis in retinal Müller cells. METHODS. Rat retinal Müller cells (rMC-1) grown in normal (N) or HG (30 mM glucose) medium for 7 days were subjected to MitoTracker Red staining to identify the mitochondrial network. Digital images of mitochondria were captured in live cells under confocal microscopy and analyzed for mitochondrial morphology changes based on form factor (FF) and aspect ratio (AR) values. Mitochondrial metabolic function was assessed by measuring oxygen consumption rate (OCR) and extracellular acidification rate (ECAR) using a bioenergetic analyzer. Cells undergoing apoptosis were identified by differential dye staining and TUNEL assay, and cytochrome c levels were assessed by Western blot analysis. RESULTS. Cells grown in HG exhibited significantly increased mitochondrial fragmentation compared to those grown in N medium (FF = 1.7 ± 0.1 vs. 2.3 ± 0.1; AR = 2.1 ± 0.1 vs. 2.5 ± 0.2; P < 0.01). OCR and ECAR were significantly reduced in cells grown in HG medium compared to those grown in N medium (steady state: 75% ± 20% of control, P < 0.02; 64% ± 22% of control, P < 0.02, respectively). These cells also exhibited a significant increase (~2-fold) in the number of apoptotic cells compared to those grown in N medium (P < 0.01), with a concomitant increase in cytochrome c levels (247% ± 94% of control, P < 0.05). CONCLUSIONS. Findings indicate that HG-induced mitochondrial morphology changes and subsequent mitochondrial dysfunction may contribute to retinal Müller cell loss associated with diabetic retinopathy.

Original languageEnglish (US)
Pages (from-to)2915-2921
Number of pages7
JournalInvestigative Ophthalmology and Visual Science
Volume58
Issue number7
DOIs
StatePublished - Jun 1 2017

Fingerprint

Diabetic Retinopathy
Glucose
Cytochromes c
Oxygen Consumption
Apoptosis
Staining and Labeling
In Situ Nick-End Labeling
Confocal Microscopy
Energy Metabolism
Mitochondria
Coloring Agents
Cell Count
Western Blotting

Keywords

  • Diabetes
  • Diabetic retinopathy
  • High glucose
  • Mitochondria
  • Retinal müller cells

ASJC Scopus subject areas

  • Ophthalmology
  • Sensory Systems
  • Cellular and Molecular Neuroscience

Cite this

Tien, Thomas ; Zhang, Joyce ; Muto, Tetsuya ; Kim, Dongjoon ; Sarthy, Vijay P ; Roy, Sayon. / High glucose induces mitochondrial dysfunction in retinal müller cells : Implications for diabetic retinopathy. In: Investigative Ophthalmology and Visual Science. 2017 ; Vol. 58, No. 7. pp. 2915-2921.
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abstract = "PURPOSE. To investigate whether high glucose (HG) induces mitochondrial dysfunction and promotes apoptosis in retinal M{\"u}ller cells. METHODS. Rat retinal M{\"u}ller cells (rMC-1) grown in normal (N) or HG (30 mM glucose) medium for 7 days were subjected to MitoTracker Red staining to identify the mitochondrial network. Digital images of mitochondria were captured in live cells under confocal microscopy and analyzed for mitochondrial morphology changes based on form factor (FF) and aspect ratio (AR) values. Mitochondrial metabolic function was assessed by measuring oxygen consumption rate (OCR) and extracellular acidification rate (ECAR) using a bioenergetic analyzer. Cells undergoing apoptosis were identified by differential dye staining and TUNEL assay, and cytochrome c levels were assessed by Western blot analysis. RESULTS. Cells grown in HG exhibited significantly increased mitochondrial fragmentation compared to those grown in N medium (FF = 1.7 ± 0.1 vs. 2.3 ± 0.1; AR = 2.1 ± 0.1 vs. 2.5 ± 0.2; P < 0.01). OCR and ECAR were significantly reduced in cells grown in HG medium compared to those grown in N medium (steady state: 75{\%} ± 20{\%} of control, P < 0.02; 64{\%} ± 22{\%} of control, P < 0.02, respectively). These cells also exhibited a significant increase (~2-fold) in the number of apoptotic cells compared to those grown in N medium (P < 0.01), with a concomitant increase in cytochrome c levels (247{\%} ± 94{\%} of control, P < 0.05). CONCLUSIONS. Findings indicate that HG-induced mitochondrial morphology changes and subsequent mitochondrial dysfunction may contribute to retinal M{\"u}ller cell loss associated with diabetic retinopathy.",
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High glucose induces mitochondrial dysfunction in retinal müller cells : Implications for diabetic retinopathy. / Tien, Thomas; Zhang, Joyce; Muto, Tetsuya; Kim, Dongjoon; Sarthy, Vijay P; Roy, Sayon.

In: Investigative Ophthalmology and Visual Science, Vol. 58, No. 7, 01.06.2017, p. 2915-2921.

Research output: Contribution to journalArticle

TY - JOUR

T1 - High glucose induces mitochondrial dysfunction in retinal müller cells

T2 - Implications for diabetic retinopathy

AU - Tien, Thomas

AU - Zhang, Joyce

AU - Muto, Tetsuya

AU - Kim, Dongjoon

AU - Sarthy, Vijay P

AU - Roy, Sayon

PY - 2017/6/1

Y1 - 2017/6/1

N2 - PURPOSE. To investigate whether high glucose (HG) induces mitochondrial dysfunction and promotes apoptosis in retinal Müller cells. METHODS. Rat retinal Müller cells (rMC-1) grown in normal (N) or HG (30 mM glucose) medium for 7 days were subjected to MitoTracker Red staining to identify the mitochondrial network. Digital images of mitochondria were captured in live cells under confocal microscopy and analyzed for mitochondrial morphology changes based on form factor (FF) and aspect ratio (AR) values. Mitochondrial metabolic function was assessed by measuring oxygen consumption rate (OCR) and extracellular acidification rate (ECAR) using a bioenergetic analyzer. Cells undergoing apoptosis were identified by differential dye staining and TUNEL assay, and cytochrome c levels were assessed by Western blot analysis. RESULTS. Cells grown in HG exhibited significantly increased mitochondrial fragmentation compared to those grown in N medium (FF = 1.7 ± 0.1 vs. 2.3 ± 0.1; AR = 2.1 ± 0.1 vs. 2.5 ± 0.2; P < 0.01). OCR and ECAR were significantly reduced in cells grown in HG medium compared to those grown in N medium (steady state: 75% ± 20% of control, P < 0.02; 64% ± 22% of control, P < 0.02, respectively). These cells also exhibited a significant increase (~2-fold) in the number of apoptotic cells compared to those grown in N medium (P < 0.01), with a concomitant increase in cytochrome c levels (247% ± 94% of control, P < 0.05). CONCLUSIONS. Findings indicate that HG-induced mitochondrial morphology changes and subsequent mitochondrial dysfunction may contribute to retinal Müller cell loss associated with diabetic retinopathy.

AB - PURPOSE. To investigate whether high glucose (HG) induces mitochondrial dysfunction and promotes apoptosis in retinal Müller cells. METHODS. Rat retinal Müller cells (rMC-1) grown in normal (N) or HG (30 mM glucose) medium for 7 days were subjected to MitoTracker Red staining to identify the mitochondrial network. Digital images of mitochondria were captured in live cells under confocal microscopy and analyzed for mitochondrial morphology changes based on form factor (FF) and aspect ratio (AR) values. Mitochondrial metabolic function was assessed by measuring oxygen consumption rate (OCR) and extracellular acidification rate (ECAR) using a bioenergetic analyzer. Cells undergoing apoptosis were identified by differential dye staining and TUNEL assay, and cytochrome c levels were assessed by Western blot analysis. RESULTS. Cells grown in HG exhibited significantly increased mitochondrial fragmentation compared to those grown in N medium (FF = 1.7 ± 0.1 vs. 2.3 ± 0.1; AR = 2.1 ± 0.1 vs. 2.5 ± 0.2; P < 0.01). OCR and ECAR were significantly reduced in cells grown in HG medium compared to those grown in N medium (steady state: 75% ± 20% of control, P < 0.02; 64% ± 22% of control, P < 0.02, respectively). These cells also exhibited a significant increase (~2-fold) in the number of apoptotic cells compared to those grown in N medium (P < 0.01), with a concomitant increase in cytochrome c levels (247% ± 94% of control, P < 0.05). CONCLUSIONS. Findings indicate that HG-induced mitochondrial morphology changes and subsequent mitochondrial dysfunction may contribute to retinal Müller cell loss associated with diabetic retinopathy.

KW - Diabetes

KW - Diabetic retinopathy

KW - High glucose

KW - Mitochondria

KW - Retinal müller cells

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DO - 10.1167/iovs.16-21355

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JO - Investigative Ophthalmology and Visual Science

JF - Investigative Ophthalmology and Visual Science

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