Abstract
Parathyroid hormone (PTH) controls two proximal tubular brush border membrane transport systems, Na+/phosphate co-transport and Na+/H+ exchange. In OK cells, a cell line with proximal tubular transport characteristics, PTH acts via kinase C and kinase A activation to inhibit Na+/phosphate co-transport [6, 8, 9, 19, 22]. In the present study, we show that PTH inhibits Na+/H+ exchange and that this effect can be mimicked by pharmacological activation of kinase A and kinase C. Ionomycin-dependent increases in cytoplasmic Ca2+ concentration do not induce inhibition of Na+/H+ exchange; PTH-dependent inhibition of Na+/H+ exchange is not prevented by ionomycin or by the intracellular Ca2+ chelator 1,2-bis(2-aminophenoxy)ethane-N,N,N′,N′-tetraacetic acid (Ca2+ clamping). Detailed dose-response curves for the different agonists, given either alone or in combination, suggest that the two regulatory cascades (kinase A and kinase C) are operating independent of each other and reach a common final target, resulting in 40–50% inhibition of Na+/H+ exchange. An analysis of intracellular pH sensitivity of Na+/H+ exchange suggests that inhibition is not related to a shift in set point, but is rather explained by a reduced V max of Na+/H+ exchange and/or reduced affinity for protons at the internal membrane surface. It is suggested that kinase A as well as kinase C can mediate PTH inhibition of renal proximal tubular Na+/H+ exchange and that the relative importance of a particular regulatory cascade is determined by the PTH-concentration-dependent rates in the liberation of diacylglycerol (phospholipase C/kinase C) and cAMP (adenylate cyclase/kinase A).
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Abbreviations
- HEPES:
-
4-(2-hydroxyethyl)-1-piperazineethane-sulphonic acid
- EDTA:
-
ethylenediaminetetraacetic acid
- FCS:
-
fetal calf serum
- PTH:
-
parathyroid hormone
- Tris:
-
2-amino-2-hydroxyme-thylpropane-1,3-diol
- BCECF:
-
2′,7′-bis(2-carboxyethyl)-5,6-carboxyfluorescein
- Pi :
-
inorganic phosphate
- pHi :
-
intracellular pH
- Cai :
-
cytosolic free Ca2+
- BAPTA:
-
1,2-bis(2-aminophenoxy)ethane-N,N,N′,N′-tetraacetic acid
- EGTA:
-
ethylene glycol-bis(β-amino-ethylether)-N,N,N′,N′-tetraacetic acid
- IP3 :
-
inositol 1,4,5-trisphos-phate
- DAG:
-
1,2-diacylglycerol
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Helmle-Kolb, C., Montrose, M.H., Stange, G. et al. Regulation of Na+/H+ exchange in opossum kidney cells by parathyroid hormone, cyclic AMP and phorbol esters. Pflügers Arch 415, 461–470 (1990). https://doi.org/10.1007/BF00373624
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DOI: https://doi.org/10.1007/BF00373624