Control of phosphate transport in flounder renal proximal tubule primary cultures

1989 ◽  
Vol 256 (4) ◽  
pp. R850-R857 ◽  
Author(s):  
A. Gupta ◽  
J. L. Renfro
Keyword(s):  
Proximal Tubule ◽  
Primary Cultures ◽  
Phosphate Transport ◽  
Renal Proximal Tubule ◽  
Pseudopleuronectes Americanus ◽  
Ussing Chambers ◽  
Long Latency ◽  
Phosphate Availability ◽  
Phosphate Fluxes ◽  
Renal Proximal Tubule Cells

Unidirectional mucosal-to-serosal (Jm----s) and serosal-to-mucosal (Js----m) transepithelial phosphate fluxes across monolayers of flounder (Pseudopleuronectes americanus) renal proximal tubule cells in primary culture were examined for effects of diacylglycerols, phorbol ester, A23187, forskolin, and extracellular phosphate availability. Tissues were cultured on floating collagen rafts and studied short circuited in Ussing chambers. Transepithelial electrical properties were continuously monitored and were unaffected by any of the treatments compared with paired controls. Under usual conditions (phosphate = 0.4 mM) tissues invariably displayed net phosphate reabsorption [Js----m = 2.3 +/- 0.52; Jm----a = 7.1 +/- 1.77; Jnet = 4.9 +/- 1.45 (SE) nmol.cm-2.h-1]. Acute elevation of bath phosphate concentration above 0.5 mM stimulated net secretion. Exposure to 100 microM 1,2-dihexanoyl-sn-glycerol stimulated net phosphate secretion within 30 min, the result of a fivefold increase in Js----m. Phorbol-12,13-didecanoate stimulated net phosphate secretion by increasing Js----m and decreasing Jm----s. The inactive diacylglycerol, 1,3-didecanoyl-rac-glycerol (100 microM), had no effect on phosphate fluxes. A23187 stimulated net phosphate secretion; Jm----s was reduced almost fourfold while Js----m was increased threefold. Forskolin (10 microM) stimulated net reabsorption more than threefold after a long latency (2 h). These data indicate that renal phosphate secretion and reabsorption may be regulated by several putative intracellular messengers. In addition, extracellular phosphate availability may modulate renal phosphate handling.

Defective PTH regulation of sodium-dependent phosphate transport in NHERF-1−/− renal proximal tubule cells and wild-type cells adapted to low-phosphate media

2005 ◽  
Vol 289 (4) ◽  
pp. F933-F938 ◽  
Author(s):  
Rochelle Cunningham ◽  
Xiaofei E ◽  
Deborah Steplock ◽  
Shirish Shenolikar ◽  
Edward J. Weinman
Keyword(s):  
Proximal Tubule ◽  
Primary Cultures ◽  
Phosphate Transport ◽  
Inhibitory Response ◽  
Renal Proximal Tubule ◽  
Wild Type ◽  
Proximal Tubule Cells ◽  
Sodium Dependent ◽  
Tubule Cells ◽  
Renal Proximal Tubule Cells

The present experiments using primary cultures from renal proximal tubule cells examine two aspects of the regulation of sodium-dependent phosphate transport and membrane sodium-dependent phosphate transporter (Npt2a) expression by parathyroid hormone (PTH). Sodium-dependent phosphate transport in proximal tubule cells from wild-type mice grown in normal-phosphate media averaged 4.4 ± 0.5 nmol·mg protein−1·10 min−1 and was inhibited by 30.5 ± 8.6% by PTH (10−7 M). This was associated with a 32.7 ± 5.2% decrease in Npt2a expression in the plasma membrane. Proximal tubule cells from Na+/H+ exchanger regulatory factor-1 (NHERF-1)−/− mice had a lower rate of phosphate transport compared with wild-type cells and a significantly reduced inhibitory response to PTH. Wild-type cells incubated in low-phosphate media for 24 h had a higher rate of phosphate transport compared with wild-type cells grown in normal-phosphate media but a significantly blunted inhibitory response to PTH. These data indicate a role for NHERF-1 in mediating the membrane retrieval of Npt2a and the subsequent inhibition of phosphate transport in renal proximal tubules. These studies also suggest that there is a blunted phosphaturic effect of PTH in cells adapted to low-phosphate media.

Primary cultures of renal proximal tubule cells derived from individuals with primary hyperoxaluria

2009 ◽  
Vol 37 (3) ◽  
pp. 127-132 ◽  
Author(s):  
Karen L. Price ◽  
Sally-Anne Hulton ◽  
William G. van’t Hoff ◽  
John R. Masters ◽  
Gill Rumsby
Keyword(s):  
Proximal Tubule ◽  
Primary Cultures ◽  
Primary Hyperoxaluria ◽  
Renal Proximal Tubule ◽  
Proximal Tubule Cells ◽  
Tubule Cells ◽  
Renal Proximal Tubule Cells

scholarly journals Signaling pathways utilized by PTH and dopamine to inhibit phosphate transport in mouse renal proximal tubule cells

2009 ◽  
Vol 296 (2) ◽  
pp. F355-F361 ◽  
Author(s):  
Rochelle Cunningham ◽  
Rajatsubhra Biswas ◽  
Marc Brazie ◽  
Deborah Steplock ◽  
Shirish Shenolikar ◽  
...  
Keyword(s):  
Proximal Tubule ◽  
Signaling Pathways ◽  
Phosphate Transport ◽  
Inhibitory Effect ◽  
Renal Proximal Tubule ◽  
Proximal Tubule Cells ◽  
Phosphate Cotransport ◽  
Tubule Cells ◽  
Renal Proximal Tubule Cells ◽  
Sodium Phosphate Cotransport

The present experiments were designed to detail factors regulating phosphate transport in cultured mouse proximal tubule cells by determining the response to parathyroid hormone (PTH), dopamine, and second messenger agonists and inhibitors. Both PTH and dopamine inhibited phosphate transport by over 30%. The inhibitory effect of PTH was completely abolished in the presence of chelerythrine, a PKC inhibitor, but not by Rp-cAMP, a PKA inhibitor. By contrast, both chelerythrine and Rp-cAMP blocked the inhibitory effect of dopamine. Chelerythrine inhibited PTH-mediated cAMP accumulation but also blocked the inhibitory effect of 8-bromo-cAMP on phosphate transport. On the other hand, Rp-cAMP had no effect on the ability of DOG, a PKC activator, to inhibit phosphate transport. PD98059, an inhibitor of MAPK, had no effect on PTH- or dopamine-mediated inhibition of sodium-phosphate cotransport. Finally, compared with 8-bromo-cAMP, 8-pCPT-2′- O-Me-cAMP, an activator of EPAC, had no effect on phosphate transport. These results outline significant differences in the signaling pathways utilized by PTH and dopamine to inhibit renal phosphate transport. Our results also suggest that activation of MAPK is not critically involved in PTH- or dopamine-mediated inhibition of phosphate transport in mouse renal proximal tubule cells in culture.

Cyclosporin A and vehicle toxicity in primary cultures of rabbit renal proximal tubule cells

1990 ◽  
Vol 183 (6) ◽  
pp. 2438
Author(s):  
P.P. Sokol ◽  
L.C. Capodagli ◽  
M. Dixon ◽  
P.D. Holohan ◽  
C.R. Ross ◽  
...  
Keyword(s):  
Proximal Tubule ◽  
Cyclosporin A ◽  
Primary Cultures ◽  
Renal Proximal Tubule ◽  
Proximal Tubule Cells ◽  
Tubule Cells ◽  
Renal Proximal Tubule Cells

Stimulation of renal sulfate secretion by metabolic acidosis requires Na+/H+ exchange induction and carbonic anhydrase

2005 ◽  
Vol 289 (1) ◽  
pp. F208-F216 ◽  
Author(s):  
Ryan M. Pelis ◽  
Susan L. Edwards ◽  
Stan C. Kunigelis ◽  
James B. Claiborne ◽  
J. Larry Renfro
Keyword(s):  
Metabolic Acidosis ◽  
Carbonic Anhydrase ◽  
Proximal Tubule ◽  
Brush Border ◽  
Primary Cultures ◽  
Acute Effect ◽  
Renal Proximal Tubule ◽  
Ussing Chambers ◽  
Renal Tubular ◽  
Nhe Isoforms

The acute effect of metabolic acidosis on SO42− secretion by the marine teleost renal proximal tubule was examined. Metabolic acidosis was mimicked in primary cultures of winter flounder renal proximal tubule epithelium (fPTCs) mounted in Ussing chambers by reducing interstitial pH to 7.1 (normally 7.7). fPTCs with metabolic acidosis secreted SO42− at a net rate that was 40% higher than in paired isohydric controls (pH 7.7 on interstitium). The stimulation was completely blocked by the carbonic anhydrase inhibitor methazolamide (100 μM). Although Na+/H+ exchange (NHE) isoforms 1, 2, and 3 were identified in fPTCs by immunoblotting, administering EIPA (20 μM) to the interstitial and luminal bath solutions had no effect on net SO42− secretion by fPTCs with a normal interstitial pH of 7.7. However, EIPA (20 μM) blocked most of the stimulation caused by acidosis when applied to the lumen but not interstitium, demonstrating that induction of brush-border NHE activity is important. In the intact flounder, serum pH dropped 0.4 pH units (pH 7.7 to 7.3, at 2–3 h) when environmental pH was lowered from 7.8 to ∼4.3. Whereas serum [SO42−] was not altered by acidosis, renal tubular SO42− secretion rate was elevated 200%. Thus metabolic acidosis strongly stimulates renal sulfate excretion most likely by a direct effect on active renal proximal tubule SO42− secretion. This stimulation appears to be dependent on inducible brush-border NHE activity.

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