title: Parathormone sensitivity and responses to protein kinases in subclones of opossum kidney cells.
abstract: Parathyroid hormone (PTH) inhibits sodium-dependent phosphate (Na(+)-Pi) transport in the renal proximal tubule and opossum kidney (OK) cells by mechanisms involving protein kinases (PK) A and C, and 20-hydroxyeicosatetraneoic acid (20-HETE). The magnitude of the effect of PKA and PKC on Na(+)-Pi transport in OK cells varies in different studies, suggesting that OK cell subclones are functionally heterogeneous despite their morphological similarity. We studied the effect of PTH and PK effectors in two separate sets of OK cells at two different time periods. Each group of cells were derived from the same stock, at passages 75-85. In one group of OK cells 20-HETE (10(-7 )M) induced a 24% decrease in Na-(32)Pi transport. Addition of PTH (10(-7) M) inhibited Pi transport by 44%. Addition of TPA (10(-8) M) resulted in a 32% decrease in Na-(32)Pi transport. Exposure of cells to the PKC inhibitor staurosporine (10(-7) M) induced a significant increase in Na-(32)Pi transport. Simultaneous addition of 20-HETE and staurosporine restored baseline Pi transport. Finally, Br-cAMP (10(-7) M) inhibited Na-(32)Pi transport by 32%. In another group of OK cells we reexamined the affect of these substances on Na-(32)Pi transport. 20-HETE (10(-7) M) induced a significant increase (30%) in Na-(32)Pi transport. PTH (10(-7) M) had no effect on Na-(32)Pi transport (P = 0.05). TPA (10(-8) M) induced a 42% increase in Na-(32)Pi transport (P < 0.01). Staurosporine (10(-7) M) induced a slight decrease in Na-(32)Pi transport (P < 0.05). Simultaneous addition of 20-HETE and staurosporine restored Na-(32)Pi transport to baseline levels. Finally, Br-cAMP (10(-7) M) inhibited Na-(32)Pi transport by 23%. We conclude that different groups OK cells have markedly different responses to regulators of Na-Pi cotransport.
authors.affiliation: Division of Nephrology, Department of Pediatrics, Health Sciences Center, Louisiana State University, New Orleans, 70118, USA. dsilve@lsuhsc.edu
authors.forename: Douglas M
authors.initials: DM
authors.lastname: Silverstein
authors.forename: Adrian
authors.initials: A
authors.lastname: Spitzer
authors.forename: Mario
authors.initials: M
authors.lastname: Barac-Nieto
mesh_terms: D015124:8-Bromo Cyclic Adenosine Monophosphate / Q000494:pharmacology; D000818:Animals; D002478:Cells, Cultured; D002999:Clone Cells; D017868:Cyclic AMP-Dependent Protein Kinases / Q000502:physiology*; D004338:Drug Combinations; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006893:Hydroxyeicosatetraenoic Acids / Q000494:pharmacology; D007668:Kidney / Q000166:cytology / Q000187:drug effects* / Q000201:enzymology*; D009893:Opossums; D010281:Parathyroid Hormone / Q000494:pharmacology*; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000502:physiology*; D050608:Sodium-Phosphate Cotransporter Proteins; D019311:Staurosporine / Q000494:pharmacology; D027981:Symporters / Q000037:antagonists & inhibitors / Q000378:metabolism; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
chemical_list: D004338:Drug Combinations; D004791:Enzyme Inhibitors; D006893:Hydroxyeicosatetraenoic Acids; D010281:Parathyroid Hormone; D050608:Sodium-Phosphate Cotransporter Proteins; D027981:Symporters; D015124:8-Bromo Cyclic Adenosine Monophosphate; C055987:20-hydroxy-5,8,11,14-eicosatetraenoic acid; D017868:Cyclic AMP-Dependent Protein Kinases; D011493:Protein Kinase C; D019311:Staurosporine; D013755:Tetradecanoylphorbol Acetate