The mycotoxin ochratoxin A deranges pH homeostasis in Madin-Darby canine kidney cells

Abstract

Ochratoxin A (OTA) is a nephrotoxin which blocks plasma membrane anion conductance in Madin-Darby canine kidney (MDCK) cells. Added to the culture medium, OTA transforms MDCK cells in a manner similar to exposure to alkaline stress. By means of video-imaging and microelectrode techniques, we investigated whether OTA (1 mumol/liter) affects intracellular pH (pHi), Cl- (Cl-i) or cell volume of MDCK cells acutely exposed to normal (pHnorm = 7.4) and alkaline (pHalk = 7.7) conditions. At pHnorm, OTA increased Cl-i by 2.6 +/- 0.4 mmol/liter (n = 14, P < 0.05) but had no effect on pHi. At pHalk, application of OTA increased Cl-i by 8.6 +/- 2.6 mmol/liter (n = 10, P < 0.05) and raised pHi by 0.11 +/- 0.03 (n = 8, P < 0.05). The Cl-/HCO3- exchange inhibitor DNDS (4,4'-dinitro-stilbene-2,2'-disulfonate; 10 mumol/liter) eliminated the OTA-induced changes of pHi and Cl-i. OTA did not affect cell volume under both pHnorm and pHalk conditions. We conclude that the OTA-induced blockade of plasma membrane anion conductance increases Cl-i without changing cell volume. The driving force of plasma membrane Cl-/HCO3- exchange dissipates, leading to a rise of pHi when cells are exposed to an acute alkaline load. Thus, OTA interferes with pHi and Cl-i homeostasis leading to morphological and functional alterations in MDCK cells.