Effects of parathyroid hormone on cytosolic calcium in renal proximal tubular primary cultures

1986 ◽  
Vol 251 (2) ◽  
pp. F188-F198 ◽  
Author(s):  
K. A. Hruska ◽  
M. Goligorsky ◽  
J. Scoble ◽  
M. Tsutsumi ◽  
S. Westbrook ◽  
...  
Keyword(s):  
Parathyroid Hormone ◽  
Cultured Cells ◽  
Primary Cultures ◽  
Cytosolic Calcium ◽  
Proximal Tubule Cells ◽  
Proximal Tubular ◽  
Tubule Cells ◽  
Renal Proximal Tubule Cells ◽  
Renal Proximal Tubular ◽  
Mitochondrial Uncouplers

The effects of parathyroid hormone on the cytoplasmic Ca2+ concentration of canine renal proximal tubule cells grown in primary culture were determined using the fluorescent Ca2+ indicator quin 2. The cultured cells exhibited responses to hormones, enzyme activities, transport functions, and morphology characteristic of the proximal convoluted tubule. Parathyroid hormone stimulated an immediate rise in cytoplasmic Ca2+, both in suspended cells and cells studied as a monolayer on Nuclepore filters. The rise in cytoplasmic Ca2+ induced by the hormone was sustained for 15-30 min, was dose dependent, and was not mimicked by cyclic AMP. Removing Ca2+ from the extracellular media markedly decreased cytoplasmic Ca2+ and abolished the effects of parathyroid hormone on cytosolic Ca2+. 8-(N,N-diethylamino)octyl-3,4,5-trimethoxybenzoate blocked the effects of the hormone on cytosolic Ca2+, but mitochondrial uncouplers failed to inhibit the effects of the hormone to increase cytoplasmic Ca2+. These studies support a role of Ca2+ in the activation of proximal renal tubule cells by parathyroid hormone.

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

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

scholarly journals Defective Parathyroid Hormone Regulation of NHE3 Activity and Phosphate Adaptation in Cultured NHERF-1-/-Renal Proximal Tubule Cells

2004 ◽  
Vol 279 (36) ◽  
pp. 37815-37821 ◽  
Author(s):  
Rochelle Cunningham ◽  
Deborah Steplock ◽  
Fengying Wang ◽  
Huijun Huang ◽  
Xiaofei E ◽  
...  
Keyword(s):  
Parathyroid Hormone ◽  
Proximal Tubule ◽  
Renal Proximal Tubule ◽  
Hormone Regulation ◽  
Proximal Tubule Cells ◽  
Tubule Cells ◽  
Renal Proximal Tubule Cells

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.

Improved culture conditions stimulate gluconeogenesis in primary cultures of renal proximal tubule cells

1995 ◽  
Vol 268 (4) ◽  
pp. C1053-C1061 ◽  
Author(s):  
G. Nowak ◽  
R. G. Schnellmann
Keyword(s):  
Proximal Tubule ◽  
Primary Cultures ◽  
Lactate Production ◽  
Glucose Production ◽  
Renal Proximal Tubule ◽  
Proximal Tubule Cells ◽  
Tubule Cells ◽  
Lactate Consumption ◽  
Renal Proximal Tubule Cells

Unlike renal proximal tubule cells (RPTC) in vivo, RPTC cultured in standard conditions are hypoxic, glycolytic, and not gluconeogenic. This study investigated the effects of glucose and lactate on glycolysis and gluconeogenesis in rabbit RPTC cultured in conditions of increased oxygen supply (Shake). Confluent Shake cultures grown in the presence of glucose exhibited increased oxygen consumption and decreased glycolysis compared with stationary (Still) cultures. Addition of 5 mM lactate to a 5 mM glucose medium decreased net glucose consumption and glucose oxidation in Shake cultures by 34 and 50%, respectively, and resulted in net lactate consumption. Addition of 5 mM lactate to a glucose-free medium resulted in a threefold increase in net glucose production (0.024 +/- 0.003 vs. 0.074 +/- 0.013 mumol.mg protein-1.day-1) in Shake cultures. Net glucose production further increased to 0.430 +/- 0.020 and 1.640 +/- 0.040 mumol.mg protein-1.day-1 when glucose reuptake was inhibited by 1 mM phloridzin or 1 mM phloridzin + 1 mM phloretin, respectively. These results show that, under conditions of improved oxygenation and in the presence of lactate and physiological levels of glucose and insulin, RPTC aerobic metabolism increases and glucose metabolism changes from glycolysis and net lactate production to gluconeogenesis and net lactate consumption.

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