RESUMO
BACKGROUND: Matrix extracellular phosphoglycoprotein (MEPE) is a putative phosphatonin that we have shown in previous studies to be phosphaturic in rats. Its site of action in the nephron remains to be confirmed. METHODS: We made micropuncture collections from late proximal convoluted tubules in anaesthetized rats to assess directly the effect of MEPE on phosphate reabsorption in the proximal tubule. RESULTS: MEPE had no effect on glomerular filtration rate or single-nephron filtration rate, but it increased phosphate excretion significantly. In animals infused with vehicle alone (time controls), no significant change was seen in either the proximal tubular fluid:plasma phosphate concentration ratio (TF/P(Pi)) or the fraction of filtered phosphate reaching the late proximal convoluted tubule (FD(Pi)); whereas in rats infused with MEPE, TF/P(Pi) increased from 0.49 ± 0.07 to 0.68 ± 0.04 (n = 22; P = 0.01) and FD(Pi) increased from 0.20 ± 0.03 to 0.33 ± 0.03 (n = 22; P < 0.01). CONCLUSIONS: The results confirm the phosphaturic effect of MEPE and indicate that much, if not all, of this effect is a result of reduced reabsorption of phosphate in the proximal convoluted tubule. This is consistent with the recent finding of MEPE-induced reductions in apically located NaPT2a in the proximal tubule.
Assuntos
Proteínas da Matriz Extracelular/fisiologia , Glicoproteínas/fisiologia , Túbulos Renais Proximais/metabolismo , Fosfatos/metabolismo , Fosfoproteínas/fisiologia , Punções , Absorção , Animais , Taxa de Filtração Glomerular , Masculino , Hormônio Paratireóideo/farmacologia , Ratos , Ratos Sprague-DawleyRESUMO
BACKGROUND: Matrix extracellular phosphoglycoprotein (MEPE), first isolated from tumour-derived tissue from a patient with oncogenic hypophosphataemia, is a putative phosphatonin that has received much less attention than fibroblast growth factor-23. To date, its effect on renal tubular phosphate reabsorption remains undefined. METHODS: A renal clearance study was performed in anaesthetized rats infused intravenously with a range of doses of MEPE. RESULTS: MEPE had no effect on glomerular filtration rate (inulin clearance) but caused rapid, dose-dependent increases in absolute and fractional phosphate excretion, wholly attributable to reduced phosphate reabsorption. At a maximal dose, MEPE increased fractional phosphate excretion more than 2-fold, whereas no change was observed in time controls. CONCLUSION: The results lend support to the hypothesis that MEPE contributes to the phosphaturia of oncogenic hypophosphataemia and of hypophosphataemic rickets.
Assuntos
Proteínas da Matriz Extracelular/fisiologia , Glicoproteínas/fisiologia , Hipofosfatemia Familiar/etiologia , Túbulos Renais/metabolismo , Fosfatos/metabolismo , Fosfoproteínas/fisiologia , Absorção , Animais , Masculino , Ratos , Ratos Sprague-DawleyRESUMO
BACKGROUND: The study of renal tubular transport requires the ability to accurately measure ion concentrations in samples taken from single tubules. Sample collection and analysis are laborious, so methods allowing determination of multiple ion species in a small volume sample are advantageous. This article describes a method for the simultaneous analysis of anions at physiologic concentrations in nanoliter volumes of tubular fluid. METHOD: The analysis is performed using capillary zone electrophoresis. Diluted samples are moved along a capillary by bulk transport and separated according to charge and size. Peaks corresponding to anions are obtained by ultraviolet (UV) detection; peak area is proportional to ion concentration. RESULTS: The anions chloride, nitrate, citrate, phosphate, and bicarbonate were separated in less than 4 minutes, and iothalamate in less than 5 minutes. Simultaneous quantitative analysis was performed for chloride, phosphate, and bicarbonate, demonstrating detection limits of 12 fmol for chloride, 12 fmol for phosphate, and 72 fmol for bicarbonate. A comparison between this method and a flow-through microfluorimeter analysis of chloride showed good agreement between the two micro-methods. Illustrative data from proximal and distal tubular fluid samples obtained by micropuncture (volume 30-70 nL) are given, as are results from urine samples. RESULTS: Results for chloride, phosphate, and bicarbonate in control material are in close agreement with the certified values, while values in tubular fluid are in accordance with previously published results. CONCLUSION: This method provides a straightforward means of analyzing multiple anions in small volume biological samples.