Effects of phorbol myristate acetate chemotactic peptide on transmembrane potentials and cytosolic free calcium in mature granulocytes evolve sequentially as the cells differentiate (original) (raw)
Related papers
Journal of leukocyte biology, 1990
Treatment of human leukemic HL-60 cells with N,N-dimethylformamide (DMF) induces them to mature until they reach granulocytoid morphology 3-6 d later. We have reported a maturation-dependent ability of these cells to respond to phorbol myristate acetate (PMA), as evaluated by membrane depolarization and by oxidative burst product formation (Newburger et al.: J. Biol. Chem. 259,3771, 1984). More recently we have attempted to develop techniques for simultaneous evaluation of these parameters during HL-60 cell maturation. Here, we compare the cytoplasmic [Ca++] and membrane potential changes elicited by the chemotactic peptide fMLP via simultaneous measurement of individual cells in a fluorescence-activated cell sorter (FACS), as done previously for mature granulocytes (Lazzari et al.: J. Biol. Chem. 261,9710, 1986). The stimulus-induced [Ca++]in changes are detected with the fluorescent probe Indo-1 and reproducibly increase in magnitude for a subpopulation of cells as the cells matur...
Biochimica et Biophysica Acta (BBA) - Molecular Basis of Disease, 2001
In mouse mammary epithelial C127 cells expressing wild-type cystic fibrosis transmembrane conductance regulator (CFTR), chloride efflux, measured with the Cl 3 -sensitive dye 6-methoxy-N-(3-sulfopropyl)quinolinium (SPQ), was stimulated by activation of protein kinase A with cyclic AMP elevating agents forskolin plus 3-isobutyl-1-methyl-xanthine (IBMX) and, to a less extent, by activation of protein kinase C with the phorbol 12-myristate 13-acetate (PMA). Conversely, bicarbonate influx, determined by intracellular alkalinization of cells incubated with the pH-sensitive dye 2P,7P-bis-(2-carboxyethyl)-5(6)-carboxyfluoresceintetraacetoxymethyl ester (BCECF-AM), was stimulated by cyclic AMP elevation, but not by PMA. Patch clamp analysis revealed that PMA activated a Cl 3 current with the typical biophysical characteristics of swelling-activated current and not of CFTR. ß
Comparison of PAF- and fMLP-induced [Ca2+]i transients in human polymorphonuclear leukocytes
Journal of Lipid Mediators and Cell Signalling, 1996
Changes of [Ca'+ Ii in human polymorphonuclear leukocytes (PMNL) were studied. PMNL suspension was activated three times every 5 min with lo-' M PAF and fMLP. Both PAF and fMLP, induced three consecutive [Ca2+li transients in PMNL suspended in medium with 1 mM Ca2+. The first Ca2+ response was a result of Ca2+ release from internal stores and the extracellular Ca" influx, while the second and third responses were completely dependent on Ca*+ influx from extracellular space. The contribution of Ca 2+ from intracellular stores to the first PAF-induced Ca2+ response was about 1.4-fold lower in comparison with the first fMLP induced Ca2+ response (27 f 1 vs 37 f 6% (p < 0.05). Previous addition of PAF enhanced 3-fold (p < 0.001) the PMNL response to fMLP while cells pretreated with fMLP failed to increase their [CaZfli after challenge with PAF. PMNL from 40% of donors did not respond to PAF in the presence of 100 nM Ca 21-However, the cells responding to PAF as the cells treated . with fMLP or cyclopiazonic acid released almost the entire Ca 2+ from intracellular stores after challenge. Subtraction of mean [Ca2+li transients in the presence of 100 nM Ca2+ from that obtained in medium with 1 mM Ca 2+ showed that, in PMNL stimulated with PAF in contrast to the cells treated with fMLP, the onset of Ca2+ influx from extracellular space precedes Ca2+ release from intracellular stores. These results suggest that PAF-induced Ca2' influx from extracellular space is at least partly independent of CaZf release from intracellular stores.
Biochemical and Biophysical Research Communications, 1984
The ability of a myeloid leukemia cell line (HL-60) to undergo membrane electrical potential changes was followed during neutrophilic differentiation induced by 2 compounds. Membrane-potential changes were induced with 12-O-tetradecanoylphorbol 13acetate (TPA) or formyl-methionyl-leucyl-phenylalanine (FMLP) and were monitored by flow cytometry. The magnitude of the membrane-potential response to TPA increased in a more uniform manner as the population of cells matured than did acquisition of mature morphology or ability to undergo the respiratory burst in response to TPA. The response to TPA and FMLP of HL-60 cells, maximally induced to differentiate by dimethylsulfoxide, closely resembled that of neutrophils. Thus, HL-60 cells may be a useful tool in the study of the relation between membrane depolarization and subsequent cellular activation.
Phorbol 12-myristate 13-acetate-mediated signalling in murine bone marrow cells
Biochemical Journal, 1995
Phorbol 12-myristate 13-acetate (PMA)-mediated signalling was investigated in relation to the ability of murine (CBA) bone marrow cells to form colonies in vitro. Treatment of marrow cells with PMA did not influence the 1,2-diacylglycerol or cyclic AMP concentrations, the intracellular Ca2+ concentration or phospholipase D activity. PMA increased particulate phospholipase A2 (PLA2) activity, lysophosphatidylcholine formation and arachidonic acid release from bone marrow cells; these effects were abolished when cells were pretreated with the putative PLA2 inhibitors heparin and mepacrine. While indomethacin and nordihydroguaiaretic acid inhibited either the cyclo-oxygenase or lipoxygenase pathway of arachidonic acid metabolism, as measured by their products prostaglandin E2 and leukotriene B4, they did not influence PMA-mediated PLA2 activation or translocation of protein kinase C (PKC) from the soluble to the particulate fraction. Treatment of cells with PMA increased the amounts of...
Journal of Clinical Investigation, 1984
We studied the effect of phorbol myristate acetate (PMA) on the plasma membrane ATPdependent calcium pump in neutrophils. Plasma membrane-enriched fractions ("podosomes") from PMAstimulated guinea pig neutrophils exhibited a twofold stimulation of ATP-dependent calcium transport when compared with control podosomes. The stimulatory effect was rapid (beginning less than 2 min after exposure to PMA) and reached maximal values within 5 min. PMA increased the maximum velocity but not the affinity of the calcium pump for Ca".
Agents and Actions, 1992
The effects of the phorbol ester, phorbol-12-myristate-13-acetate (PMA), and of the calcium ionophore A23187 on DNA synthesis in murine quiescent and mitogen-stimulated lymphocytes have been examined. Neither PMA nor A23187 had any mitogen effect on their own on quiescent lymphocytes. However, stimulation of cells sequentially with A23187 and then PMA resulted in a proliferative response in proportion to the duration of the exposure to A23187, and the sustained simultaneous presence of both agents was necessary for maximum proliferation. On the other hand, while short incubations with A23187 potentiated mitogen-induced DNA synthesis, prolonged exposure inhibited it. Furthermore, on lymphocytes stimulated with two T cell mitogens, the effects of A23187 and PMA depended on the proliferation-inducing mitogens and the responsiveness level induced by them. Therefore, while PMA and short pretreatments with A23187 had no effect on high intensity mitogenic responses, low intensity responses were significantly enhanced. These results demonstrate differential effects of A23187 and PMA on DNA synthesis that should be useful in studies on the mechanisms of activation of lymphocyte proliferation.
Biochimica Et Biophysica Acta - Biomembranes, 1986
The electrophysiological properties of the membrane of mouse peritoneal macrophage polykaryons are studied. Slow hyperpolarizations can be elicited by iontophoretic injections of either Ca 2+ or Sr 2+ into the cytoplasm. The effect of both cations is identical, since: (1) it is invariably triggered by the cation injection, (2) the amplitude is dependent on the K + gradient, (3) quinine blocks reversibly the response to both cation injections. Mg 2+, Ba 2+ and Mn 2+ did not elicit responses when injected into the cytoplasm. Ca 2+ induced slow hyperpolarizations were reversibly blocked by the addition of Ba 2+ to the external saline, but were not affected by the presence of external tetraethylammonium chloride. Cells maintained in saline containing high concentrations of Ca 2+, Sr 2+ or Mn 2+ exhibited sustained hyperpolarizations. Quinine blocked the hyperpolarization induced by high Ca 2+ or Sr 2+, but was ineffective for the case of Mn 2+. Cells hyperpolarized by external Mn 2+ frequently exhibited nonlinear, voltage-current characteristics. Similar patterns could also be observed in a small fraction (less than 10%) of the cells in control conditions. Current-induced shifts between two stable membrane potentials were seen either in high Ca 2+ or normal medium. The great variability of the responses described for this phagocytic membrane is discussed. The evidence supports the assumption that Ca 2+ and Sr 2+ can induce transient or persistent hyperpolarized states by activating a potassium permeability. External Mn 2 ÷ may act in part by reducing impalement-related current leakage from the phagocytic membrane.
Journal of Membrane Biology, 1992
The membrane electric effects of N,N′-dicyclohexylcarbodiimide (DCCD) and vanadate were studied in murine erythroleukemia cells (MELC), comparing the patch-clamp technique and the accumulation ratio (AR exp) of [3H]-tetraphenylphosphonium (TPP+). Electrophysiological measurements showed that both these inhibitors produce, at micromolar concentrations, a 20–30 mV hyperpolarization of resting potential (Δψ p ) of MELC, which is abolished when the electrochemical equilibrium potential of K+ (E K) is brought close to zero. DCCD and vanadate turned out to have distinct targets on the plasma membrane of MELC (an H+ pump and the Na+,K+-ATPase, respectively). Measurements of AR exp showed that: (i) patch-clamp measurements of Δψ p were equivalent to those based on ARexp of antimycin-pretreated cells (AR ANT); (ii) DCCD produced a strong increase in AR ANT, that was antagonized by carbonyl cyanide p-trifluoromethoxyphenyl-hydrazone (FCCP) and diethylstilbestrol (DES); (iii) vanadate determined a marked increase in AR ANT that was insensitive to FCCP, but antagonized by ouabain; (iv) incubation in high K+ medium (HK) brought ARANT to 1.0 in the controls, but did not lower this ratio below 3.0 in the presence of DCCD or vanadate; (v) the total amount of TPP+ taken up by the cells was in any case water extractable by a freezing and thawing procedure. On the whole, our data indicate that DCCD and vanadate hyperpolarize the MELC by increasing the K+ conductance and, at the same time, enhance the TPP+ binding, probably by changing the electrostatic potential profile of the plasma membrane. These effects seem to involve functional modifications of the target pumps, apparently related to the ion-occluding state of these enzymes. We gratefully acknowledge the revision of the English by Dr. R. Manning of the Department of Biological Sciences, University of Durham, Durham, UK. A. Arcangeli was supported by a fellowship from the Associazione Italiana contro le Leucemie (AIL); A. Becchetti and M.R. Del Bene were supported by fellowships from the Italian Association for Cancer Research (AIRC). This work was supported by grants from the AIRC, from the Consiglio Nazionale delle Ricerche (CNR) (Special project “Ion channels”) and from the Ministero Pubblica Istruzione.
Biochimica et Biophysica Acta (BBA) - Biomembranes, 1978
The membrane potential of the Ehrlich ascites tumor cell was shown to be influenced by its amino acid content and the activity of the Na ÷ : K ÷ pump. The membrane potential (monitored by the fluorescent dye, 3,3'-dipropylthiodicarbocyanine iodide) varied with the size of the endogenous amino acid pool and with the concentration of accumulated 2-aminoisobutyrate. When cellular amino acid content was high, the cells were hyperpolarized; as the pool declined in size, the cells were depolarized. The hyperpolarization seen with cellular amino acid required cellular Na ÷ but not cellular ATP. Na ÷ efflux was more rapid from cells containing 2-aminoisobutyrate than from cells low in internal amino acids. These observations indicate that the hyperpolarization recorded in cells with high cellular amino acid content resulted from the electrogenic co-efflux of Na ÷ and amino acids. Cellular ATP levels were found to decline rapidly in the presence of the dye and hence the influence of the pump was seen only if glucose was added to the cells. When the cells contained normal Na ÷ (approx. 30 mM), the Na + : K ÷ pump was shown to have little effect on the membrane potential (the addition of ouabain had little effect on the potential). When cellular Na ÷ was raised to 60 mM, the activity of the pump changed the membrane potential from the range-25 to-30 mV to-44 to-63 mV. This hyperpolarization required external K ÷ and was inhibited by ouabain.