It is the long term goal of this research to characterize all the significant transport mechanisms in corneal epithelium and endothelium which both produce electric current and contribute to fluid transport by these cells and to determine the specific role of these transporters in corneal deturgescence and in the maintenance of corneal transparency.
Six specific aims are proposed: 1) Measure the currents produced by the sodium pump, the sodium-bicarbonate cotransporter, the sodium-calcium exchanger, and the sodium-alanine transporter in single isolated cells of corneal epithelium and endothelium. 2) Study the bicarbonate stimulation of the potassium selective channel from rabbit corneal endothelial. 3) Characterize the properties of individual ion channels in corneal endothelium and corneal epithelium. 4) Utilize patch clamp techniques to measure electrical properties of endothelial cells in whole corneas and in the isolated descemet's- endothelial monolayer. 5) Determine the effect of culture on selected transporters in cornea epithelium and endothelium. 6) Study the properties and gating control of gap junctions between corneal epithelium and corneal endothelium. Patch clamp techniques will be used to characterize the detailed properties of single K+, cation, and anion channels in these cells. Three different techniques will be used to measure whole currents and mechanisms for controlling these currents will be investigated. Optical techniques with and without specific fluorescent probes will be used to measure cell volume and its control in corneal epithelium and endothelium and to measure Na+, K+, Ca++, and pH in these cells. Both optical and electrophysiological techniques will be used to measure gap junction properties and the control of connexon permeability.
| Rae, J L; Levis, R A (2004) Fabrication of patch pipets. Curr Protoc Neurosci Chapter 6:Unit 6.3 |
| Mathias, Richard T; Rae, James L (2004) The lens: local transport and global transparency. Exp Eye Res 78:689-98 |
| Ou, Yijun; Strege, Peter; Miller, Steven M et al. (2003) Syntrophin gamma 2 regulates SCN5A gating by a PDZ domain-mediated interaction. J Biol Chem 278:1915-23 |
| Rae, James L; Levis, Richard A (2002) Single-cell electroporation. Pflugers Arch 443:664-70 |
| Rae, J L; Shepard, A R (2000) Kir2.1 Potassium channels and corneal epithelia. Curr Eye Res 20:144-52 |
| Rae, J L; Shepard, A R (2000) Kv3.3 potassium channels in lens epithelium and corneal endothelium. Exp Eye Res 70:339-48 |
| Rich, A; Farrugia, G; Rae, J L (1999) Effects of melatonin on ionic currents in cultured ocular tissues. Am J Physiol 276:C923-9 |
| Shepard, A R; Rae, J L (1999) Electrically silent potassium channel subunits from human lens epithelium. Am J Physiol 277:C412-24 |
| Shepard, A R; Rae, J L (1999) ""Microprep"" method for rapidly isolating plasmid DNAs for restriction enzyme analysis. Biotechniques 26:868-70 |
| Shepard, A R; Rae, J L (1998) Ion transporters and receptors in cDNA libraries from lens and cornea epithelia. Curr Eye Res 17:708-19 |
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