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Manik Goel, Adam E. Sienkiewicz, Richard K. Lee, Sanjoy K. Bhattacharya; Concomitant Expression of Cochlin and TREK-1 Modulates Trabecular Meshwork Cell Function. Invest. Ophthalmol. Vis. Sci. 2011;52(14):4610.
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To determine the role of cochlin and TREK-1 on the functional and physiological behavior of trabecular meshwork cells.
Primary human trabecular meshwork (TM) cells were cultured from cadaveric corneo-scleral rims obtained from the Bascom Palmer Eye Institute Eye Bank. TM cells were co-transfected with TREK-1 and cochlin or TREK-1 and RPE65 (control) expression vectors. Transfected cells were subjected to capillary tube collagen gel expansion assay (5000 cells/assay) and fluorescein dye flow was measured across polyvinylidene fluoride membranes (PVDF) membrane with confluent tri-layer of TM cells in an Ussing-type chamber. Capillaries were digitally imaged at 0, 24 and 48 hours post transfection and analyzed using NIH ImageJ (v.1.43u) software. Fluorescein flow across the PVDF membrane was determined by analyzing the sampled solution using the spectrophotometer. All experiments were repeated at least five times and statistically analyzed.
Capillary tubes with cells transfected with TREK-1+cochlin showed 10.3±2.1 percent gel expansion, in contrast to 3.82±0.9 percent gel contraction in capillary tubes with TREK-1+RPE65 control transfected cells. TM cells transfected with TREK-1+cochlin showed significantly greater levels of fluorescein flow across the PVDF membrane than those transfected with TREK-1+RPE65.
Cochlin and TREK-1 co-expression results in opening up of more spaces between the TM cells as indicated by collagen gel expansion and increased fluorescein flow across PVDF membrane. Cochlin may play a part in regulating aqueous outflow in the TM in conjunction with TREK-1.
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