PMID- 36552890
OWN - NLM
STAT- MEDLINE
DCOM- 20221226
LR  - 20230115
IS  - 2073-4409 (Electronic)
IS  - 2073-4409 (Linking)
VI  - 11
IP  - 24
DP  - 2022 Dec 19
TI  - Relative Importance of Different Elements of Mitochondrial Oxidative 
      Phosphorylation in Maintaining the Barrier Integrity of Retinal Endothelial 
      Cells: Implications for Vascular-Associated Retinal Diseases.
LID - 10.3390/cells11244128 [doi]
LID - 4128
AB  - PURPOSE: Mitochondrial dysfunction is central to breaking the barrier integrity 
      of retinal endothelial cells (RECs) in various blinding eye diseases such as 
      diabetic retinopathy and retinopathy of prematurity. Therefore, we aimed to 
      investigate the role of different mitochondrial constituents, specifically those 
      of oxidative phosphorylation (OxPhos), in maintaining the barrier function of 
      RECs. METHODS: Electric cell-substrate impedance sensing (ECIS) technology was 
      used to assess in real time the role of different mitochondrial components in the 
      total impedance (Z) of human RECs (HRECs) and its components: capacitance (C) and 
      the total resistance (R). HRECs were treated with specific mitochondrial 
      inhibitors that target different steps in OxPhos: rotenone for complex I, 
      oligomycin for complex V (ATP synthase), and FCCP for uncoupling OxPhos. 
      Furthermore, data were modeled to investigate the effects of these inhibitors on 
      the three parameters that govern the total resistance of cells: Cell-cell 
      interactions (R(b)), cell-matrix interactions (alpha), and cell membrane permeability 
      (Cm). RESULTS: Rotenone (1 microM) produced the greatest reduction in Z, followed by 
      FCCP (1 microM), whereas no reduction in Z was observed after oligomycin (1 microM) 
      treatment. We then further deconvoluted the effects of these inhibitors on the 
      R(b), alpha, and C(m) parameters. Rotenone (1 microM) completely abolished the resistance 
      contribution of R(b), as the R(b) became zero immediately after the treatment. 
      Secondly, FCCP (1 microM) eliminated the resistance contribution of R(b) only after 
      2.5 h and increased C(m) without a significant effect on alpha. Lastly, of all the 
      inhibitors used, oligomycin had the lowest impact on R(b), as evidenced by the 
      fact that this value became similar to that of the control group at the end of 
      the experiment without noticeable effects on C(m) or alpha. CONCLUSION: Our study 
      demonstrates the differential roles of complex I, complex V, and OxPhos coupling 
      in maintaining the barrier functionality of HRECs. We specifically showed that 
      complex I is the most important component in regulating HREC barrier integrity. 
      These observed differences are significant since they could serve as the basis 
      for future pharmacological and gene expression studies aiming to improve the 
      activity of complex I and thereby provide avenues for therapeutic modalities in 
      endothelial-associated retinal diseases.
FAU - Eltanani, Shaimaa
AU  - Eltanani S
AD  - Department of Ophthalmology, Visual, and Anatomical Sciences, Wayne State 
      University, Detroit, MI 48201, USA.
FAU - Yumnamcha, Thangal
AU  - Yumnamcha T
AD  - Department of Ophthalmology, Visual, and Anatomical Sciences, Wayne State 
      University, Detroit, MI 48201, USA.
FAU - Gregory, Andrew
AU  - Gregory A
AD  - Department of Ophthalmology, Visual, and Anatomical Sciences, Wayne State 
      University, Detroit, MI 48201, USA.
FAU - Elshal, Mahmoud
AU  - Elshal M
AUID- ORCID: 0000-0002-9528-7483
AD  - Department of Ophthalmology, Visual, and Anatomical Sciences, Wayne State 
      University, Detroit, MI 48201, USA.
AD  - Department of Pharmacology and Toxicology, Faculty of Pharmacy, Mansoura 
      University, Mansoura 35516, Egypt.
AD  - Department of Pharmacology, Wayne State University, Detroit, MI 48201, USA.
FAU - Shawky, Mohamed
AU  - Shawky M
AD  - Department of Ophthalmology, Visual, and Anatomical Sciences, Wayne State 
      University, Detroit, MI 48201, USA.
AD  - Department of Biochemistry, Faculty of Pharmacy, Horus University, New Damietta 
      34518, Egypt.
FAU - Ibrahim, Ahmed S
AU  - Ibrahim AS
AUID- ORCID: 0000-0001-8480-6252
AD  - Department of Ophthalmology, Visual, and Anatomical Sciences, Wayne State 
      University, Detroit, MI 48201, USA.
AD  - Department of Pharmacology, Wayne State University, Detroit, MI 48201, USA.
AD  - Department of Biochemistry, Faculty of Pharmacy, Mansoura University, Mansoura 
      35516, Egypt.
LA  - eng
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, Non-U.S. Gov't
DEP - 20221219
PL  - Switzerland
TA  - Cells
JT  - Cells
JID - 101600052
RN  - 03L9OT429T (Rotenone)
RN  - 370-86-5 (Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone)
RN  - 0 (Oligomycins)
SB  - IM
MH  - Infant, Newborn
MH  - Humans
MH  - *Oxidative Phosphorylation
MH  - Rotenone/pharmacology
MH  - Endothelial Cells/metabolism
MH  - Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone/metabolism/pharmacology
MH  - Mitochondria/metabolism
MH  - *Diabetic Retinopathy/metabolism
MH  - Oligomycins/pharmacology
PMC - PMC9776835
OTO - NOTNLM
OT  - ECIS modeling
OT  - FCCP
OT  - OxPhos
OT  - Rb resistance
OT  - barrier integrity
OT  - capacitance
OT  - human retinal endothelial cells (HRECs)
OT  - impedance
OT  - oligomycin
OT  - oxidative phosphorylation
OT  - rotenone
OT  - alpha resistance
COIS- The authors declare no conflict of interest.
EDAT- 2022/12/24 06:00
MHDA- 2022/12/27 06:00
PMCR- 2022/12/19
CRDT- 2022/12/23 01:18
PHST- 2022/10/22 00:00 [received]
PHST- 2022/12/06 00:00 [revised]
PHST- 2022/12/13 00:00 [accepted]
PHST- 2022/12/23 01:18 [entrez]
PHST- 2022/12/24 06:00 [pubmed]
PHST- 2022/12/27 06:00 [medline]
PHST- 2022/12/19 00:00 [pmc-release]
AID - cells11244128 [pii]
AID - cells-11-04128 [pii]
AID - 10.3390/cells11244128 [doi]
PST - epublish
SO  - Cells. 2022 Dec 19;11(24):4128. doi: 10.3390/cells11244128.