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RNA sequencing uncovers alterations in corneal endothelial metabolism, pump and barrier functions of Slc4a11 KO mice.
- Source :
-
Experimental eye research [Exp Eye Res] 2022 Jan; Vol. 214, pp. 108884. Date of Electronic Publication: 2021 Dec 03. - Publication Year :
- 2022
-
Abstract
- 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.<br /> (Copyright © 2021 Elsevier Ltd. All rights reserved.)
- Subjects :
- Animals
Blotting, Western
Corneal Edema metabolism
Fluorescent Antibody Technique, Indirect
Glutamine metabolism
Glycolysis
Mice
Mice, Knockout
Oxidative Stress
Oxygen Consumption physiology
Real-Time Polymerase Chain Reaction
Sequence Analysis, RNA
Anion Transport Proteins genetics
Corneal Edema genetics
Endothelium, Corneal metabolism
Monocarboxylic Acid Transporters metabolism
Muscle Proteins metabolism
Sodium-Potassium-Exchanging ATPase metabolism
Symporters genetics
Symporters metabolism
Subjects
Details
- Language :
- English
- ISSN :
- 1096-0007
- Volume :
- 214
- Database :
- MEDLINE
- Journal :
- Experimental eye research
- Publication Type :
- Academic Journal
- Accession number :
- 34871568
- Full Text :
- https://doi.org/10.1016/j.exer.2021.108884