Journal
AMERICAN JOURNAL OF PHYSIOLOGY-RENAL PHYSIOLOGY
Volume 292, Issue 2, Pages F674-F681Publisher
AMER PHYSIOLOGICAL SOC
DOI: 10.1152/ajprenal.00272.2006
Keywords
proteinuria; renal; nephropathy; permeability; slit diaphragm
Categories
Funding
- Medical Research Council [G0500053] Funding Source: researchfish
- MRC [G0500053] Funding Source: UKRI
- Medical Research Council [G0500053] Funding Source: Medline
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Glomerular podocytes are critical regulators of glomerular permeability via the slit diaphragm and may play a role in cleaning the glomerular filter. Whether podocytes are able to endocytose proteins is uncertain. We studied protein endocytosis in conditionally immortalized mouse and human podocytes using FITC-albumin by direct quantitative assay and by fluorescence microscopy and electron microscopy in mouse podocytes. Furthermore, in vivo uptake was studied in human, rat, and mouse podocytes. Both mouse and human podocytes displayed specific one-site binding for FITC-albumin with K-d of 0.91 or 0.44 mg/ml and B-max of 3.15 or 0.81 mu g/mg cell protein, respectively. In addition, they showed avid endocytosis of FITC-albumin with Km of 9.48 or 4.5 mg/ml and V-max of 474.3 or 97.4 mu g(.)mg cell protein(-1) (.)h(-1), respectively. Immunoglobulin and transferrin were inefficient competitors of this process, indicating some specificity for albumin. Accumulation of endocytosed albumin could be demonstrated in intracellular vesicles by fluorescence confocal microscopy and electron microscopy. Endocytosis was sensitive to pretreatment with simvastatin. In vivo accumulation of albumin was found in all three species but was most pronounced in the rat. We conclude that podocytes are able to endocytose protein in a statin-sensitive manner. This function is likely to be highly significant in health and disease. In addition, protein endocytosis by podocytes may represent a useful, measurable phenotypic characteristic against which potentially injurious or beneficial interventions can be assessed.
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