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Am J Physiol Renal Physiol (January 29, 2002). doi:10.1152/ajprenal.00340.2001
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Articles in PresS, published online ahead of print January 28, 2002
Am J Physiol Renal Physiol, 10.1152/ajprenal.00340.2001
Submitted on November 13, 2001
Accepted on January 25, 2002

Expression and function of sodium transporters in two opossum kidney cell clonal sublines

Pedro Gomes1, Jing Xu2, Paula Serrao1, Sofia Doria3, Pedro A Jose2, and Patricio Soares-da-Silva1*

1 Institute of Pharmacology & Therapeutics, Faculty of Medicine, Porto, Portugal
2 Departments of Pediatrics and Physiology & Biophysics, Georgetown University Medical Center, Washington, DC, USA
3 Department Genetics, Faculty of Medicine, Porto, Portugal

The present study describes characteristic features of two clonal subpopulations of opossum kidney (OK) cells (OKLC and OKHC) that are functionally different but morphologically identical. The most impressive differences between OKHC and OKLC cells are that the former over-expressed Na+-K+ ATPase and type 3 Na+/H+ exchanger, accompanied by an increased Na+-K+ ATPase activity (57.6±5.6 vs 30.0±0.1 nmol Pi/mg protein/min), increased ability to translocate Na+ from the apical to the basolateral surface, and increased Na+-dependent pHi recovery (0.254±0.016 vs 0.094±0.011 pH units/s). Vmax values (in pH units/s) for Na+-dependent pHi recovery in OKHC cells (0.00521±0.0004) were twice (P<0.05) those in OKLC (0.00202±0.0001), with similar Km values (in mM) for Na+ (OKLC, 21.0±5.5; OKHC, 14.0±5.6). In addition, we measured the activities of transporters (organic ions, {alpha}-methyl-D-glucoside, L-type amino acids and Na+) and enzymes (adenylyl cyclase, aromatic L-amino acid decarboxylase and catechol-O-methyltransferase). The cells were also characterized morphologically by optical and scanning electron microscopy and karyotyped. It is suggested that OKLC and OKHC cells constitute an interesting cell model for the study of renal epithelial physiology and pathophysiology, namely hypertension.




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