The goal of this study is to understand how the corneal endothelial fluid "pump" works. The "pump" maintains corneal hydration and transparency. When the "pump" fails due to trauma, inflammation, ageing, or corneal dystrophy, corneal edema ensues, transparency is lost and vision is significantly degraded. The usual therapy is transplantation, which is not without significant compromises and complications. Knowing how the "pump" works is one approach to developing medical therapies that could delay or supplant the need for transplantation. As the population ages and the prevalence of endothelial dysfunction increases, demand for endothelial therapy will also increase. Ion transport is a key feature of the "pump". Up to now the "pump" has been modeled as a classic ion secretory mechanism that is bicarbonate dependent and carbonic anhydrase sensitive. Previous studies and our work in the preceding period however, indicate that alternate models need to be investigated. Our overall hypothesis is that the corneal endothelium is a lactate removal "pump". Because the cornea is very glycolytic and must remove the end product, which is lactic acid, we propose that the transcellular flux of lactate is coupled to water movement;that lactate flux is via monocarboxylic acid transporters (MCTs);and that the buffering action of HCO3-, CA activity, membrane pHi regulators &HCO3- transporters act in concert to facilitate the flux of lactate. By buffering the protons transported by the MCTs, lactate:H+ &water flux is maximized by preventing reductions in the driving force for continued lactate:H+ transport. Using multiple in vitro &in vivo complementary approaches this will be tested in three aims.
Aim 1 will investigate the role of buffering capacity on water and lactate fluxes. The flux ratio, i.e. mMoles lactate/ul water can be estimated to determine the tonicity of this osmotic coupling. If water coupled to lactate is significant, we expect isotonic (300 mEq/L) transport.
Aim 2 will investigate the role of primary and secondary active transport (Na+,K+ ATPase,1Na+:2HCO3- cotransport, and Na+/H+ exchange) in facilitating lactate flux across the endothelium by determining the change in lactate flux ratio when these transporters are disturbed.
Aim 3 will investigate the role of MCTs in facilitated lactate transport. Using pharmacological inhibitors and shRNA approaches in vivo we predict that inhibition of MCTs will have significant effects on corneal hydration. If the hypothesis is correct, we will have a more complete model of endothelial function that will allow further development of diagnostic and medical therapies or engineering of endothelial-like cells with the requisite transport properties.

Public Health Relevance

The goal of this study is to understand how the corneal endothelial fluid pump works to keep the cornea transparent. Experiments are designed to test the hypothesis that the endothelium transports water from stroma to anterior chamber by facilitating the flux of lactic acid.

Agency
National Institute of Health (NIH)
Institute
National Eye Institute (NEI)
Type
Research Project (R01)
Project #
5R01EY008834-22
Application #
8444410
Study Section
Anterior Eye Disease Study Section (AED)
Program Officer
Mckie, George Ann
Project Start
1991-07-01
Project End
2017-02-28
Budget Start
2013-03-01
Budget End
2014-02-28
Support Year
22
Fiscal Year
2013
Total Cost
$370,500
Indirect Cost
$133,000
Name
Indiana University Bloomington
Department
None
Type
Schools of Optometry/Ophthalmol
DUNS #
006046700
City
Bloomington
State
IN
Country
United States
Zip Code
47401
Robertson, Danielle M; Alexander, Larry J; Bonanno, Joseph A et al. (2014) Cornea and ocular surface disease: application of cutting-edge optometric research. Optom Vis Sci 91:S3-16
Li, Shimin; Nguyen, Tracy T; Bonanno, Joseph A (2014) CD147 required for corneal endothelial lactate transport. Invest Ophthalmol Vis Sci 55:4673-81
Jalimarada, Supriya S; Ogando, Diego G; Vithana, Eranga N et al. (2013) Ion transport function of SLC4A11 in corneal endothelium. Invest Ophthalmol Vis Sci 54:4330-40
Ogando, Diego G; Jalimarada, Supriya S; Zhang, Wenlin et al. (2013) SLC4A11 is an EIPA-sensitive Na(+) permeable pHi regulator. Am J Physiol Cell Physiol 305:C716-27
Bonanno, Joseph A (2012) Molecular mechanisms underlying the corneal endothelial pump. Exp Eye Res 95:2-7
Nguyen, Tracy T; Bonanno, Joseph A (2012) Lactate-Hýýý transport is a significant component of the in vivo corneal endothelial pump. Invest Ophthalmol Vis Sci 53:2020-9
Li, Shimin; Allen, Kah Tan; Bonanno, Joseph A (2011) Soluble adenylyl cyclase mediates bicarbonate-dependent corneal endothelial cell protection. Am J Physiol Cell Physiol 300:C368-74
Liu, Cailing; Ogando, Diego; Bonanno, Joseph A (2011) SOD2 contributes to anti-oxidative capacity in rabbit corneal endothelial cells. Mol Vis 17:2473-81
Nguyen, Tracy T; Bonanno, Joseph A (2011) Bicarbonate, NBCe1, NHE, and carbonic anhydrase activity enhance lactate-H+ transport in bovine corneal endothelium. Invest Ophthalmol Vis Sci 52:8086-93
Liu, Cailing; Cheng, Qiang; Nguyen, Tracy et al. (2010) Knockdown of NBCe1 in vivo compromises the corneal endothelial pump. Invest Ophthalmol Vis Sci 51:5190-7

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