PMID- 34871568
OWN - NLM
STAT- MEDLINE
DCOM- 20220211
LR  - 20230102
IS  - 1096-0007 (Electronic)
IS  - 0014-4835 (Print)
IS  - 0014-4835 (Linking)
VI  - 214
DP  - 2022 Jan
TI  - RNA sequencing uncovers alterations in corneal endothelial metabolism, pump and 
      barrier functions of Slc4a11 KO mice.
PG  - 108884
LID - S0014-4835(21)00450-4 [pii]
LID - 10.1016/j.exer.2021.108884 [doi]
AB  - Slc4a11 KO mice show significant corneal edema, altered endothelial morphology, 
      and mitochondrial ROS at an early age without a decrease in endothelial cell 
      density. We examined the differential gene expression profile between wild type 
      (WT) and KO with the goal of finding pathways related to corneal endothelial 
      metabolic, pump and barrier function that can explain the corneal edema. Freshly 
      dissected Corneal Endothelium-Descemet's Membrane (CEDM) and cultured Mouse 
      Corneal Endothelial Cells (MCEC) were obtained from WT and Slc4a11 KO mice. RNA 
      sequencing Ingenuity Pathway Analysis (IPA) predicted activation, inhibition or 
      differential regulation of several pathways. QPCR and Western analysis validated 
      downregulation of Glycolytic enzymes, Mitochondrial complex components and Ion 
      transporters. Functional testing revealed decreases in endothelial lactate 
      production, Extracellular Acidification Rate (ECAR), glutaminolysis, and Oxygen 
      Consumption Rate (OCR) of KO CEDM in the presence of Glutamine (Gln) that was not 
      compensated by fatty acid oxidation. Stromal lactate was significantly elevated 
      in KO along with decreased expression of MCT1 and MCT4 lactate transporters in 
      endothelial cells. ATP levels were 2x higher in KO CEDM, concomitant with a 
      3-fold decrease in Na-K-ATPase activity and reduced basolateral membrane 
      localization. Genes for cholesterol biosynthesis, glutathione metabolism and 
      tight and adherens junctions were elevated. Alteration of tight junction 
      structure and cortical cytoskeleton is evident in KO corneal endothelium with a 
      significant increase in trans-endothelial fluorescein permeability. We conclude 
      that Slc4a11 KO induces a coordinated decrease in glycolysis, glutaminolysis, 
      lactate transport and Na-K-ATPase activity. These changes together with an 
      altered barrier function cause an accumulation of stromal lactate in Slc4a11 KO 
      mice leading to chronic corneal edema.
CI  - Copyright (c) 2021 Elsevier Ltd. All rights reserved.
FAU - Ogando, Diego G
AU  - Ogando DG
AD  - Vision Science Program, School of Optometry, Indiana University, Bloomington, IN, 
      47405, USA. Electronic address: digogand@indiana.edu.
FAU - Bonanno, Joseph A
AU  - Bonanno JA
AD  - Vision Science Program, School of Optometry, Indiana University, Bloomington, IN, 
      47405, USA. Electronic address: jbonanno@indiana.edu.
LA  - eng
GR  - R01 EY008834/EY/NEI NIH HHS/United States
GR  - R01 EY031321/EY/NEI NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
DEP - 20211203
PL  - England
TA  - Exp Eye Res
JT  - Experimental eye research
JID - 0370707
RN  - 0 (Anion Transport Proteins)
RN  - 0 (Monocarboxylic Acid Transporters)
RN  - 0 (Muscle Proteins)
RN  - 0 (Slc16a4 protein, mouse)
RN  - 0 (Slc4a11 protein, mouse)
RN  - 0 (Symporters)
RN  - 0 (monocarboxylate transport protein 1)
RN  - 0RH81L854J (Glutamine)
RN  - EC 7.2.2.13 (Sodium-Potassium-Exchanging ATPase)
SB  - IM
MH  - Animals
MH  - Anion Transport Proteins/*genetics
MH  - Blotting, Western
MH  - Corneal Edema/*genetics/metabolism
MH  - Endothelium, Corneal/*metabolism
MH  - Fluorescent Antibody Technique, Indirect
MH  - Glutamine/metabolism
MH  - Glycolysis
MH  - Mice
MH  - Mice, Knockout
MH  - Monocarboxylic Acid Transporters/*metabolism
MH  - Muscle Proteins/*metabolism
MH  - Oxidative Stress
MH  - Oxygen Consumption/physiology
MH  - Real-Time Polymerase Chain Reaction
MH  - Sequence Analysis, RNA
MH  - Sodium-Potassium-Exchanging ATPase/*metabolism
MH  - Symporters/*genetics/*metabolism
PMC - PMC8792362
MID - NIHMS1762864
OTO - NOTNLM
OT  - Barrier function
OT  - Corneal endothelial dystrophy
OT  - Lactate transporters
OT  - Metabolism
OT  - Na-K-ATPase
OT  - Oxidative stress
OT  - RNA sequencing
EDAT- 2021/12/07 06:00
MHDA- 2022/02/12 06:00
PMCR- 2023/01/01
CRDT- 2021/12/06 20:12
PHST- 2021/09/27 00:00 [received]
PHST- 2021/11/17 00:00 [revised]
PHST- 2021/11/30 00:00 [accepted]
PHST- 2021/12/07 06:00 [pubmed]
PHST- 2022/02/12 06:00 [medline]
PHST- 2021/12/06 20:12 [entrez]
PHST- 2023/01/01 00:00 [pmc-release]
AID - S0014-4835(21)00450-4 [pii]
AID - 10.1016/j.exer.2021.108884 [doi]
PST - ppublish
SO  - Exp Eye Res. 2022 Jan;214:108884. doi: 10.1016/j.exer.2021.108884. Epub 2021 Dec 
      3.