PURPOSE: To determine if albumin, the major protein component of the aqueous humor, passes into the lens in vivo. METHODS: Rat albumin was covalently-labeled with Alexa 488 fluorophore, purified by gel permeation chromatography, then injected into the aqueous chamber of living rats. At 5 min postinjection, lenses were removed and analyzed by HPLC gel permeation chromatography, confocal microscopy, and immunogold electron microscopy. RESULTS: At 5 min postinjection, HPLC analysis detected measurable amounts of Alexa-labeled albumin in the lens. The results were confirmed by confocal microscopy, which showed passage into epithelial and cortical fiber cells. Immunogold electron microscopy using antibody to the Alexa fluorophore demonstrated intracellular location of the Alexa-albumin complex. CONCLUSIONS: In vivo, significant amounts of albumin pass from the aqueous chamber into cells of the lens, consistent with a possible physiological role for this process involving passage of metabolites into the lens.
PURPOSE: To determine if albumin, the major protein component of the aqueous humor, passes into the lens in vivo. METHODS:Rat albumin was covalently-labeled with Alexa 488 fluorophore, purified by gel permeation chromatography, then injected into the aqueous chamber of living rats. At 5 min postinjection, lenses were removed and analyzed by HPLC gel permeation chromatography, confocal microscopy, and immunogold electron microscopy. RESULTS: At 5 min postinjection, HPLC analysis detected measurable amounts of Alexa-labeled albumin in the lens. The results were confirmed by confocal microscopy, which showed passage into epithelial and cortical fiber cells. Immunogold electron microscopy using antibody to the Alexa fluorophore demonstrated intracellular location of the Alexa-albumin complex. CONCLUSIONS: In vivo, significant amounts of albumin pass from the aqueous chamber into cells of the lens, consistent with a possible physiological role for this process involving passage of metabolites into the lens.
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