Fluorescence dilution technique for measurement of albumin reflection coefficient in isolated glomeruli
| Autor: | Fan Fan, Mukut Sharma, Richard J. Roman, Carlos Schreck, Jan M. Williams, Daniel A. Beard, Jin Zhang, Eric A. Roman, Virginia J. Savin, Chun Cheng Andy Chen, Takashi Hirata |
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| Rok vydání: | 2015 |
| Předmět: |
Oncotic pressure
medicine.medical_specialty Protamine sulfate Physiology Kidney Glomerulus Indicator Dilution Techniques Glomerulus (kidney) Fluorescence Permeability Streptozocin Diabetes Mellitus Experimental Rats Sprague-Dawley Diabetic nephropathy Albumins Internal medicine medicine Animals Diabetic Nephropathies Kidney Rats Inbred Dahl Chromatography Proteinuria Chemistry Albumin Articles medicine.disease Mice Inbred C57BL medicine.anatomical_structure Endocrinology Permeability (electromagnetism) medicine.symptom medicine.drug |
| Zdroj: | American Journal of Physiology-Renal Physiology. 309:F1049-F1059 |
| ISSN: | 1522-1466 1931-857X |
| DOI: | 10.1152/ajprenal.00311.2015 |
| Popis: | This study describes a high-throughput fluorescence dilution technique to measure the albumin reflection coefficient (σAlb) of isolated glomeruli. Rats were injected with FITC-dextran 250 (75 mg/kg), and the glomeruli were isolated in a 6% BSA solution. Changes in the fluorescence of the glomerulus due to water influx in response to an imposed oncotic gradient was used to determine σAlb. Adjustment of the albumin concentration of the bath from 6 to 5, 4, 3, and 2% produced a 10, 25, 35, and 50% decrease in the fluorescence of the glomeruli. Pretreatment of glomeruli with protamine sulfate (2 mg/ml) or TGF-β1 (10 ng/ml) decreased σAlbfrom 1 to 0.54 and 0.48, respectively. Water and solute movement were modeled using Kedem-Katchalsky equations, and the measured responses closely fit the predicted behavior, indicating that loss of albumin by solvent drag or diffusion is negligible compared with the movement of water. We also found that σAlbwas reduced by 17% in fawn hooded hypertensive rats, 33% in hypertensive Dahl salt-sensitive (SS) rats, 26% in streptozotocin-treated diabetic Dahl SS rats, and 21% in 6-mo old type II diabetic nephropathy rats relative to control Sprague-Dawley rats. The changes in glomerular permeability to albumin were correlated with the degree of proteinuria in these strains. These findings indicate that the fluorescence dilution technique can be used to measure σAlbin populations of isolated glomeruli and provides a means to assess the development of glomerular injury in hypertensive and diabetic models. |
| Databáze: | OpenAIRE |
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