Sphingosine inhibits phorbol 12-myristate 13-acetate, but not serum-induced, activation of Na+/H+ exchange in mammalian cells

R. J. Gillies, R. Martinez, J. M. Sneider, Patricia B Hoyer

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Abstract

Addition of serum to quiescent mammalian cells in culture initiates a series of events which culminates in DNA replication and cell division. One of the earliest events in this sequence of events is activation of Na+/H+ exchange, which can result in an increase in intracellular pH (pH(in)). The regulation of this change in activity is not known. Since treatment of 3T3 cells with activators of protein kinase C (kinase C) can result in an increased pH(in), it has been hypothesized that serum stimulation of kinase C is responsible for activation of Na+/H+ exchange. Recently, sphingolipids have been discovered to inhibit kinase C both in vitro and in vivo. Therefore, we undertook the present study to ask whether or not inhibition of kinase C using sphingolipids prevents mitogen-induced alkalinization in 3T3 cells. Our results indicate that activators of kinase C stimulate Na+/H+ exchange in normal human fibroblasts (BoGi), but not in mouse embryo (3T3) cells. Addition of serum to BoGi cells, on top of saturating doses of phorbol 12-myristate 13-acetate (PMA), results in a further cytoplasmic alkalinization. Furthermore, sphingosine prevents the PMA-induced increase in pH(in) in BoGi cells, and phorphorylation of an 80 kDa protein in 3T3 cells, but not the serum-induced alkalinization in either BoGi or 3T3 cells. These data indicate that activation of kinase C does not participate in the physiological activation of Na+/H+ exchange in human fibroblasts or mouse embryo cells by serum.

Original languageEnglish (US)
Pages (from-to)125-130
Number of pages6
JournalJournal of Cellular Physiology
Volume139
Issue number1
StatePublished - 1989

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3T3 Cells
Sphingosine
Ion exchange
Acetates
Phosphotransferases
Chemical activation
Cells
Serum
Sphingolipids
Fibroblasts
Embryonic Structures
DNA Replication
Mitogens
Cell culture
Cell Division
Cell Culture Techniques
phorbol-12-myristate
DNA
Proteins

ASJC Scopus subject areas

  • Cell Biology
  • Clinical Biochemistry
  • Physiology

Cite this

Sphingosine inhibits phorbol 12-myristate 13-acetate, but not serum-induced, activation of Na+/H+ exchange in mammalian cells. / Gillies, R. J.; Martinez, R.; Sneider, J. M.; Hoyer, Patricia B.

In: Journal of Cellular Physiology, Vol. 139, No. 1, 1989, p. 125-130.

Research output: Contribution to journalArticle

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