Biomedical subjects

P H Peters

Publications and source records attributed to P H Peters.

9 recordsLinked to original sources

PubMed1998

Epidermal growth factor (EGF) receptor density controls mitogenic activation of normal rat kidney (NRK) cells by EGF.

Normal rat kidney (NRK) fibroblasts are immortalized cells that are strictly dependent on externally added growth factors for proliferation. When cultured in the presence of epidermal growth factor (EGF) as the only growth stimulating hormone, these cells have a normal phenotype and undergo density-dependent growth inhibition. It has been postulated that this density-arrest results from a decrease of EGF receptor levels below a threshold level which makes these cells unresponsive to stimulation by EGF. In the present study, we show that NRK cells, made quiescent by serum-deprivation at submaximum density, are mitogenically still responsive to EGF, but show enhanced mitogenic stimulation after 8 hr pre-treatment with either transforming growth factor beta (TGF beta) or retinoic acid (RA), while prostaglandin F2 alpha (PGF2 alpha) and bradykinin (BK) enhance the mitogenic stimulation by EGF only slightly under these conditions. Addition of TGF beta or RA results in an increase of both 125I-EGF-binding capacity and EGF receptor mRNA levels. Using flow cytometric analysis, we show that pre-treatment with TGF beta or RA increases the percentage of cells entering the cell cycle as a function of time. Furthermore, pre-treatment of the cells with TGF beta or RA increases the rate of mitogen-activated protein kinase (MAPK) phosphorylation by EGF. PGF2 alpha and BK also increase EGF receptor levels, but only with delayed kinetics. These results show that already in serum-deprived quiescent NRK cells, EGF receptor levels limit EGF-induced mitogenic stimulation. This observation provides further evidence for the regulating role of the EGF receptor in density-dependent growth control of NRK cells.

Animals↗

PubMed1997

Synchronized calcium spiking resulting from spontaneous calcium action potentials in monolayers of NRK fibroblasts.

The correlation between the intracellular Ca2+ concentration ([Ca2+]i) and membrane potential in monolayers of density-arrested normal rat kidney (NRK) fibroblasts was investigated. Using the fluorescent probe Fura-2, spontaneous repetitive spike-like increases in [Ca2+]i (Ca2+ spikes) were observed that were synchronised throughout the entire monolayer. Ca2+ spikes disappeared in Ca(2+)-free solutions and could be blocked by the L-type Ca2+ channel antagonist felodipine. Simultaneous measurements of [Ca2+]i and membrane potential showed that these Ca2+ spikes were paralleled by depolarisations of the plasma membrane. Using patch clamp measurements, action potential-like depolarisations consisting of a fast spike depolarisation followed by a plateau phase were seen with similar kinetics as the Ca2+ spikes. The action potentials could be blocked by L-type Ca2+ channel blockers and were dependent on extracellular Ca2+. The plateau phase was predominantly determined by a Cl- conductance and was dependent on intracellular Ca2+. The presence of voltage-dependent L-type Ca2+ channels in NRK cells was confirmed by patch clamp measurements in single cells. It is concluded that monolayers of density-arrested NRK fibroblasts exhibit spontaneous Ca2+ action potentials leading to synchronised Ca2+ spiking. This excitability of monolayers of fibroblasts may represent a novel Ca2+ signaling pathway in electrically coupled fibroblasts, cells that were hitherto considered to be inexcitable.

Action Potentials↗

PubMed1994

Bradykinin-induced growth inhibition of normal rat kidney (NRK) cells is paralleled by a decrease in epidermal-growth-factor receptor expression.

Normal rat kidney fibroblasts, grown to density arrest in the presence of epidermal growth factor (EGF), can be induced to undergo phenotypic transformation by treatment with transforming growth factor beta or retinoic acid. Here we show that bradykinin blocks this growth-stimulus-induced loss of density-dependent growth arrest by a specific receptor-mediated mechanism. The effects of bradykinin are specific, and are not mimicked by other phosphoinositide-mobilizing agents such as prostaglandin F2 alpha. Northern-blot analysis and receptor-binding studies demonstrate that bradykinin also inhibits the retinoic acid-induced increase in EGF receptor levels in these cells. These studies provide additional evidence that EGF receptor levels modulate EGF-induced expression of the transformed phenotype in these cells.

Animals↗

PubMed1993

Cadmium ion is a non-competitive inhibitor of red cell Ca(2+)-ATPase activity.

In the presence as well as in the absence of calmodulin, Cd2+ inhibits the human erythrocyte plasma membrane Ca(2+)-ATPase activity non-competitively with Ki = 2 nM, whereas ATP-dependent Ca(2+)-transport across the red cell membrane was found to be inhibited competitively by Cd2+ (Verbost, P.M., Flik, G., Pang, P.K.T., Lock, R.A.C. and Wendelaar Bonga, S.E. (1989) J. Biol. Chem. 264, 5613-5615). In this study it will be argued that Cd2+ also inhibits Ca(2+)-transport non-competitively, and that the discrepancy with previous conclusions most probably relies on use of an incorrect computer program that calculates the free concentrations of Ca2+ and Cd2+ at the experimental conditions applied for measurement of Ca2+ uptake.

Cadmium↗

PubMed1987

Effect of ruthenium red upon Ca2+ and Mn2+ uptake in Saccharomyces cerevisiae. Comparison with the effect of La3+.

The initial rate of both Ca2+ and Mn2+ uptake is inhibited by ruthenium red to about the same extent as by equivalent concentrations of La3+. The inhibition of Ca2+ uptake, however, is relieved during further incubation with ruthenium red. On preincubating the cells with ruthenium red even a stimulation of divalent cation uptake can be found. Relieve of the inhibition of divalent cation uptake is accompanied by K+ efflux. Both ruthenium red and La3+ displace Ca2+ very effectively from binding sites at the cell surface. The inhibition of initial Ca2+ uptake is accompanied by a reduction in the binding of Ca2+.

Calcium↗

PubMed1987

Changes in 45Ca and 109Cd uptake, membrane potential and cell pH in Saccharomyces cerevisiae provoked by Cd2+.

The effect of Cd2+ poisoning of Saccharomyces cerevisiae on 45Ca, 109Cd and [14C]tetraphenylphosphonium (TPP) uptake and cell pH was examined. At Cd2+ concentrations that produced substantial K+ efflux the rates of uptake of 45Ca, 109Cd and [14C]TPP increased progressively during incubation of the cells with Cd2+, and the cell pH was lowered concomitantly. The initial rates of uptake of the divalent cations and of TPP were increased in cells pre-loaded with Cd2+, which shows that stimulation of the ion fluxes was exerted by the Cd2+ that accumulated in the cells. The distribution ratio of TPP between cells and medium, however, was decreased by Cd2+. Although hyperpolarization of the cell membrane by Cd2+ cannot be excluded, it is argued that Cd2+ primarily stimulated divalent cation uptake by increasing the cation permeability of the cell membrane allowing the cations to enter the cells more easily.

Cadmium↗

PubMed1986

A study of the mechanism by which inhibitors of the plasmamembrane ATPase enhance uptake of divalent cations in yeast.

The enhancement of divalent cation uptake in yeast provoked by the membrane ATPase inhibitors trifluoperazine, miconazole, compound 48/80, ethidium, DIO-9 and calmidazolium should be ascribed to an increase in cation permeability of the yeast rather than to hyperpolarisation of the yeast cell membrane. For trifluoperazine and miconazole it is unequivocally shown that the cells are hyperpolarized though for miconazole only transiently. Whether the other drugs also hyperpolarize the yeast cells is uncertain. The apparent hyperpolarisation caused by trifluoperazine and miconazole may be attributed to a specific increase in the K+ permeability of the yeast plasmamembrane evoked by these compounds.

Adenosine Triphosphatases↗

PubMed1981

Factors affecting the inhibition of yeast plasma membrane ATPase by vanadate.

Inhibition of yeast plasma membrane ATPase by vanadate occurs only if either Mg2+ or MgATP2- is bound to the enzyme. The dissociation constant of the complex of vanadate and inhibitory sites is 0.14-0.20 microM in the presence of optimal concentrations of Mg2+ and of the order of 1 microM if the enzyme is saturated with MgATP2-. The dissociation constants of Mg2+ and MgATP2- for the sites involved are 0.4 and 0.62-0.73 mM, respectively, at pH 7. KCl does not increase the affinity of vanadate to the inhibitory sites as was found with (Na+ + K+)-ATPase. On the other hand, the effect of Mg2+ upon vanadate binding is similar to that upon (Na+ + K+)-ATPase, and the corresponding affinity constants of Mg2+ and vanadate for the two enzymes are of the same order of magnitude.

Adenosine Triphosphatases↗

PubMed1977

Effect of the medium pH and the cell pH upon the kinetical parameters of phosphate uptake by yeast.

1. Both the maximum rate of phosphate uptake and the Km depend upon the pH of the medium in a complex way. 2. The effect of medium pH upon the maximum rate of uptake is mainly indirect and is correlated with changes in cell pH. 3. The Km is affected by the medium pH both directly via an apparent competitive inhibition by hydroxyl anions and indirectly in a similar way as the maximum rate of uptake.

Culture Media↗