July 2018
Volume 59, Issue 9
Open Access
ARVO Annual Meeting Abstract  |   July 2018
Ceramide-induced Mitochondrial Damage in Diabetic Retinopathy
Author Affiliations & Notes
  • Yan Levitsky
    Physiology/Chemistry, Michigan State University, East Lansing, Michigan, United States
    DO/PhD - Physician Scientist Training Program, Michigan State University, East Lansing, Michigan, United States
  • Sandra S Hammer
    Physiology, Michigan State University, East Lansing, Michigan, United States
  • Todd Lydic
    Molecular Metabolism and Disease - Mass Spectrometry Core, Michigan State University, East Lansing, Michigan, United States
    Physiology, Michigan State University, East Lansing, Michigan, United States
  • Artem Muchnik
    Chemistry, Michigan State University, East Lansing, Michigan, United States
  • Anand Saripalli
    Chemistry, Michigan State University, East Lansing, Michigan, United States
  • Philip Kirschner
    Physiology, Michigan State University, East Lansing, Michigan, United States
    Heinrich-Heine-Universität Düsseldorf, Düsseldorf, Germany
  • David Pegouske
    Physiology/Chemistry, Michigan State University, East Lansing, Michigan, United States
  • Denis Proshlyakov
    Chemistry, Michigan State University, East Lansing, Michigan, United States
  • Julia V Busik
    Physiology, Michigan State University, East Lansing, Michigan, United States
  • Footnotes
    Commercial Relationships   Yan Levitsky, None; Sandra Hammer, None; Todd Lydic, None; Artem Muchnik, None; Anand Saripalli, None; Philip Kirschner, None; David Pegouske, None; Denis Proshlyakov, None; Julia Busik, None
  • Footnotes
    Support  NIH R01 EY016077-08
Investigative Ophthalmology & Visual Science July 2018, Vol.59, 6071. doi:
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      Yan Levitsky, Sandra S Hammer, Todd Lydic, Artem Muchnik, Anand Saripalli, Philip Kirschner, David Pegouske, Denis Proshlyakov, Julia V Busik; Ceramide-induced Mitochondrial Damage in Diabetic Retinopathy. Invest. Ophthalmol. Vis. Sci. 2018;59(9):6071.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose : Increase in acid sphingomyelinase (ASM) activity and ceramide production were shown to promote diabetes-induced pro-inflammatory and pro-apoptotic changes in the retina and retinal pigment epithelial (RPE) cells. Mitochondria were recently shown to contain sphingolipids, as well as enzymes of sphingolipid pathway, including ASM. This study addressed the role of diabetes-induced increase in ASM and ceramide in mitochondrial damage in the retina and RPE cells.

Methods : Retinal mitochondria from control, STZ-induced diabetic and ASM-/- rodent models were isolated by differential centrifugation. Sphingolipids were analyzed by electrospray ionization high resolution mass spectrometry. Human (H)RPE cells were isolated from control and diabetic donors. Mitochondrial morphology and ceramide localization were measured by fluorescence microscopy. mRNA was measured by qPCR. Respiration was measured using a novel custom designed microrespirometer.

Results : Relative mitochondrial ceramide (mCer) abundance increased in short term (7 wks) diabetic rat retina (38 ± 4%) compared to control (26.5 ± 6.5%, P<0.05, n=3). Mitochondrial sphingomyelin (mSM) decreased in diabetic retina (62 ± 1%) compared to control (72.5 ± 7%, P<0.05, n=3). Long term (36 wks) diabetic (n=4) rat retina showed significant increases in short chain (C16:0, C18:0) mCer compared to control (n=3, P<0.05). Wildtype and ASM-/- mouse retinas were used to determine if mCer changes are due to ASM. ASM-/- retina showed increased mSM (85.0 ± 6.7%) compared to wildtype (77.0 ± 2.8%) and decreased mCer (11.5 ± 3.6%) compared to wildtype (17.6 ± 3.6%, P<0.05, n=3). We determined if similar changes occur in RPE layer, a mitochondrial rich cell population missing from retina preparations. HRPE isolated from diabetic donor retinas showed increased IL1ß, IL6 and ASM expression and a 2- fold increase in Cer compared to control (P<0.05, n=3). Average mitochondrial length decreased in diabetic HRPE (1.2 ± 0.57 μm) compared to control (3.4 ± 0.78 μm, P<0.05, n=3). Whole cell respiratory control ratio was decreased greater than 2-fold in diabetic HRPE compared to control.

Conclusions : Diabetes-induced elevation of ASM expression and activity leads to increased ceramide abundance in mitochondria isolated from diabetic retina and RPE cells. This increase is associated with upregulation of inflammatory markers and mitochondrial dysfunction in diabetic retina and RPE cells.

This is an abstract that was submitted for the 2018 ARVO Annual Meeting, held in Honolulu, Hawaii, April 29 - May 3, 2018.

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