Growth‐dependent subcellular redistribution of protein kinase C in cultured porcine aortic endothelial cells

Uratsuji, Yuko; DiCorleto, Paul E.

doi:10.1002/jcp.1041360306

Cited by 30 publications

(21 citation statements)

References 53 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…However, except for protein kinase C, in bovine EC, all other isoforms were predominantly located in the cytosol, supporting earlier findings (Uratsuji and Di Corleto, 1988). The distribution of protein kinase C activity in these cells (as determined by histone phosphorylation) correlated well with that of the immunoreactivity (i.e.…”

Section: Discussionsupporting

confidence: 85%

Modulation of endothelial autacoid release by protein kinase C: Feedback inhition or non‐specific attenuation of receptor‐dependent cell activation?

Hecker

Lückhoff

Busse

1993

Journal Cellular Physiology

View full text Add to dashboard Cite

Receptor-mediated elevations of intracellular Ca2+ in endothelial cells may be controlled by a negative feedback mechanism through activation of protein kinase C (PKC). To test this hypothesis, we studied the effects of an activation or inhibition of PKC on the release of nitric oxide (NO) and prostacyclin (PGI2) from cultured bovine and porcine aortic endothelial cells (EC). Preincubation with the PKC activators phorbol-12-myristate-13-acetate (PMA) (3-300 nM) or 1-oleyl-2-acetyl-glycerol (OAG) (30 microM) significantly attenuated the release of NO and PGI2 from EC stimulated with bradykinin (0.3-30 nM), whereas phorbol-12,13-didecanoate (PDD) (30-300 nM), which does not activate PKC, had no effect. UCN-01 (10 nM), a specific PKC inhibitor, significantly augmented the bradykinin-stimulated release of NO from EC. These effects were correlated with a reduced (PMA) or enhanced (UCN-01) elevation of intracellular Ca2+ in response to bradykinin in both types of EC. Neither the PKC activators nor the inhibitor had any effect on resting intracellular Ca2+ or basal endothelial autacoid release. Several isoforms of PKC (namely PKC alpha, PKC delta, PKC epsilon, and PKC zeta) were detected in bovine, human, and porcine EC by immunoblotting analysis with isotype-specific anti-PKC antibodies, which, except PKC epsilon, were predominantly located in the cytosol. Incubation of bovine EC with PMA elicited a significant increase in membrane-bound PKC alpha immunoreactivity, whereas there was no translocation of PKC alpha from the cytosolic to the membrane fraction with bradykinin. As determined by histone phosphorylation, PKC activity was similarly reduced in the cytosol, but increased in the membrane fraction of bovine EC exposed to PMA, whereas bradykinin had no significant effect. These findings indicate that endothelial autacoid release can be modulated by activators and inhibitors of PKC. However, stimulation of EC with bradykinin does not lead to a detectable activation of PKC, suggesting that PKC does not exert a negative feedback in the signal transduction pathway of this receptor-dependent agonist.

show abstract

Section: Discussionsupporting

confidence: 85%

Modulation of endothelial autacoid release by protein kinase C: Feedback inhition or non‐specific attenuation of receptor‐dependent cell activation?

Hecker

Lückhoff

Busse

1993

Journal Cellular Physiology

View full text Add to dashboard Cite

show abstract

“…Despite a 24-hour withdrawal from serum, the cells were not quiescent as previously observed by other authors using a similar protocol.14 Under control conditions, 29+8 nuclei out of 100 were labeled (mean+SD, 22 experiments, range, 17-36). ATP (10-100 ,uM) increased this labeling index to 42±6% (mean±SD, 19 experiments, range, 34-57%), which represents a 44% increase ( Figure 2).…”

Section: Mitogenic Assaysupporting

confidence: 54%

Effects of adenine nucleotides on the proliferation of aortic endothelial cells.

Daele¹,

Coevorden²,

Roger³

et al. 1992

Circulation Research

View full text Add to dashboard Cite

The effects of adenine nucleotides and adenosine on DNA synthesis and cell growth have been studied in bovine aortic endothelial cells (BAECs). ATP produced a small but significant (+44%) increase of the fraction of BAECs whose nuclei are labeled by [3H]thymidine. This mitogenic effect was mimicked by ADP, the phosphorothioate analogues ATPyS and ADP,BS, and the nonhydrolyzable analogue adenosine 5'-(,6,y-imido)triphosphate (APPNP), whereas adenosine 5'-(a3,-methylene)triphosphate (APCPP), a selective agonist of P2x-purinoceptors, had no effect at 10 ,uM and a small one at 100 ,iM; this profile is consistent with the involvement of P2y-receptors. Adenosine induced a mitogenic response of a magnitude similar to that of ATP. This effect was not reproduced by R-phenylisopropyl adenosine, by 5'-Nethylcarboxamide adenosine, or by 2',5'-dideoxyadenosine, selective ligands of the A1-and A2-receptors and the P site, respectively, nor was it inhibited by 8-phenyltheophylline, an antagonist of both A1-and A2-receptors. The mechanism of this adenosine action thus remains unclear. ATP and ATPyS did not enhance the proliferation of BAECs cultured in the presence of fetal calf serum concentrations ranging from 0.5% to 10%. They inhibited the growthpromoting effect of basic fibroblast growth factor; among the various nucleotides tested, APCPP was the least effective to reproduce the action of ATP, suggesting the possible involvement of P2y-receptors. In conclusion, the action of ATP on the proliferation of BAECs is complex: an increase in the fraction of cells synthesizing DNA, no effect on the cell proliferation in the presence of serum, and inhibition of the growth-promoting effect of basic fibroblast growth factor. (Circulation Research 1992;70:82-90) A TP induces a rapid release of endotheliumderived relaxing factor (nitric oxide) and prostacyclin (prostaglandin 12) from vascular endothelial cells.' These actions, which are mediated by P2y-receptors, are obtained in a range of ATP concentrations (1-100 ,uM) comparable to those obtained in plasma after platelet degranulation. They might thus play an important role in the interaction between platelets and the endothelium.' In endothelial cells, as in other cells, P2y-receptors are coupled to a phospholipase C that hydrolyzes phosphatidylinositol bisphosphate.12 As a consequence of this initial event, ATP induces in aortic endothelial cells several biochemical responses that are often associated with a mitogenic action: enhanced phosphorylation of 27-kd proteins,3-5 stimulation of phosphati-

show abstract

“…However, the majority of PKC activity in non-dividing cultured porcine aortic endotheiial ceiis is found in the cytoplasm, whereas in vigorously proliferating cells, >lore than half of the total activity is found in the memurane fraction [34]. This agrees with our results in immunoperoxidase stainings where no translocation was seen after IFN-1/ treatment.…”

Section: Discussionsupporting

confidence: 91%

Protein kinase C subtypes in endothelial cells

Mattila

1991

FEBS Letters

View full text Add to dashboard Cite

Activation of protein kinase C (PKC) has been linked to the regulation of class II expression on endothelial cells by interferon‐γ (IFN‐γ). PKC subtypes in endothelial cells were analyzed using three different approaches, the immunoperoxidase staining of native and IFN‐γ stimulated cells cultured on chamber slides as well as immuno‐ and Northern blotting. All approaches revealed that of the conventional subtypes. α is the predominant form of PKC in endothelial cells. Even though IFN‐γ is able to induce PKC translocation to particulate fractions, no translocation was detected in histological stainings. Western blot studies as well as mRNA studies revealed that IFN‐γ is unable to increase the total amount of PKC in endothelial cells.

show abstract

Growth‐dependent subcellular redistribution of protein kinase C in cultured porcine aortic endothelial cells

Cited by 30 publications

References 53 publications

Modulation of endothelial autacoid release by protein kinase C: Feedback inhition or non‐specific attenuation of receptor‐dependent cell activation?

Modulation of endothelial autacoid release by protein kinase C: Feedback inhition or non‐specific attenuation of receptor‐dependent cell activation?

Effects of adenine nucleotides on the proliferation of aortic endothelial cells.

Protein kinase C subtypes in endothelial cells

Contact Info

Product

Resources

About