Stable intracellular acidification upon polyamine depletion induced by α-difluoromethylornithine or N1, N12-bis(ethyl)spermine in L1210 leukaemia cells

R. Poulin, A. E. Pegg

Research output: Contribution to journalArticlepeer-review

10 Scopus citations

Abstract

Polyamines play major roles in ionic and osmotic regulation, but their exact involvement in specific ion transport processes is poorly defined. Treatment of L1210 mouse leukaemia cells with either 5mM α-difluoromethylornithine (DEMO), a suicide substrate of ornithine decarboxylase, or 25 μM N1, N12-bis(ethyl)spermine (BE-3-4-3), a dysfunctional polyamine analogue, caused a stable decrease in intracellular pH (pH,) by 0.1-0.4 unit from steady-state control values between 7.4 and 7.6, as measured either by partition of a weak acid or with a fluorescent pH-sensitive probe. This effect was not related to cell growth status or differences in metabolic acid generation, and was observed in either the presence or absence of HCO31. Exogenous spermidine (10-25 μM) or putrescine (25-50 μM) fully reversed DFMO- or BE-3-4-3-induced acidification within 2 and 8 h respectively. Recovery of pH(i) in L1210 cells after a nigericin- or NH4+-mediated acid load in HCO3- -free buffers was mediated by Na+/H+ antiporter activity, in addition to a minor Na+-independent and amiloride-insensitive pathway. Decreased steady-state pH(i) was maintained in polyamine-depleted L1210 cells after recovery from acid stress. Moreover, the pH(i)-dependence of the rate of Na+-dependent H+- extrusion after an acid stress was altered by DFMO and BE-3-4-3, resulting in a set-point which was lower by 0.25-0.30 pH unit in polyamine depleted cells. On the other hand, neither the rate nor the magnitude of Na+/H+-exchanger-mediated alkalinization induced by hypertonic shock was decreased by polyamine depletion. Thus polyamine depletion induces a persistent defect in pH, homeostasis which is due, at least in part, to a stable decrease in the pH(i) set-point of the Na+/H+ exchanger.

Original languageEnglish (US)
Pages (from-to)749-756
Number of pages8
JournalBiochemical Journal
Volume312
Issue number3
DOIs
StatePublished - 1995

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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