Involvement of PKCdelta and PKD in pulmonary microvascular endothelial cell hyperpermeability.
The involvement of PKC, the isoforms of which are categorized into three subtypes: conventional (alpha, betaI, betaII, and gamma), novel [delta, epsilon, eta, and mu (also known as PKD), theta], and atypical (zeta and iota/lambda), in the regulation of endothelial monolayer integrity is well documented. However, isoform activity varies among different cell types. Our goal was to reveal isoform-specific PKC activity in the microvascular endothelium in response to phorbol 12-myristate 13-acetate (PMA) and diacylglycerol (DAG). Isoform activity was demonstrated by cytosol-to-membrane translocation after PMA treatment and phosphorylation of the myristoylated alanine-rich C kinase substrate (MARCKS) protein after PMA and DAG treatment. Specific isoforms were inhibited by using both antisense oligonucleotides and pharmacological agents. The data showed partial cytosol-to-membrane translocation of isoforms alpha, betaI, and epsilon and complete translocation of PKCdelta and PKD in response to PMA. Furthermore, antisense treatment and pharmacological studies indicated that the novel isoform PKCdelta and PKD are both required for PMA- and DAG-induced MARCKS phosphorylation and hyperpermeability in pulmonary microvascular endothelial cells, whereas isoforms alpha, betaI, and epsilon were dispensable with regard to these same phenomena.
D000818:Animals; D001692:Biological Transport / Q000187:drug effects; D002199:Capillary Permeability / Q000187:drug effects / Q000502:physiology*; D002478:Cells, Cultured; D004075:Diglycerides / Q000494:pharmacology; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D005959:Glucosidases; D047908:Intracellular Signaling Peptides and Proteins; D008565:Membrane Proteins; D008833:Microcirculation; D000076250:Myristoylated Alanine-Rich C Kinase Substrate; D016376:Oligonucleotides, Antisense / Q000494:pharmacology; D010750:Phosphoproteins / Q000378:metabolism; D010766:Phosphorylation / Q000187:drug effects; D011493:Protein Kinase C / Q000502:physiology*; D051745:Protein Kinase C-delta; D011652:Pulmonary Circulation; D051381:Rats; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
D004075:Diglycerides; D047908:Intracellular Signaling Peptides and Proteins; C000619712:Marcks protein, rat; D008565:Membrane Proteins; D016376:Oligonucleotides, Antisense; D010750:Phosphoproteins; C512948:Prkcsh protein, rat; D000076250:Myristoylated Alanine-Rich C Kinase Substrate; C497260:Prkcd protein, rat; C089212:protein kinase D; D011493:Protein Kinase C; D051745:Protein Kinase C-delta; D005959:Glucosidases; D013755:Tetradecanoylphorbol Acetate