ABSTRACT
Endothelial cells in growing tumors express activated Akt, which when modeled by transgenic endothelial expression of myrAkt1 was sufficient to recapitulate the abnormal structural and functional features of tumor blood vessels in nontumor tissues. Sustained endothelial Akt activation caused increased blood vessel size and generalized edema from chronic vascular permeability, while acute permeability in response to VEGF-A was unaffected. These changes were reversible, demonstrating an ongoing requirement for Akt signaling for the maintenance of these phenotypes. Furthermore, rapamycin inhibited endothelial Akt signaling, vascular changes from myrAkt1, tumor growth, and tumor vascular permeability. Akt signaling in the tumor vascular stroma was sensitive to rapamycin, suggesting that rapamycin may affect tumor growth in part by acting as a vascular Akt inhibitor.
Subject(s)
Endothelial Cells/pathology , Endothelium, Vascular/pathology , Neoplasms/blood supply , Neovascularization, Pathologic/metabolism , Proto-Oncogene Proteins c-akt/metabolism , Sirolimus/pharmacology , Animals , Capillary Permeability , Cells, Cultured , Edema/metabolism , Endothelial Cells/drug effects , Endothelium, Vascular/drug effects , Humans , Mice , Mice, Transgenic , Proto-Oncogene Proteins c-akt/genetics , Rats , Signal Transduction , Vascular Endothelial Growth Factor A/physiologyABSTRACT
Stimulation of platelets with strong agonists results in centralization of cytoplasmic organelles and secretion of granules. These observations have led to the supposition that cytoskeletal contraction facilitates granule release by promoting the interaction of granules with one another and with membranes of the open canalicular system. Yet, the influence of the actin cytoskeleton in controlling the membrane fusion events that mediate granule secretion remains largely unknown. To evaluate the role of the actin cytoskeleton in platelet granule secretion, we have assessed the effects of latrunculin A and cytochalasin E on granule secretion. Exposure of platelets to low concentrations of these reagents resulted in acceleration and augmentation of agonist-induced alpha-granule secretion with comparatively modest effects on dense granule secretion. In contrast, exposure of platelets to high concentrations of latrunculin A inhibited agonist-induced alpha-granule secretion but stimulated dense granule secretion. Incubation of permeabilized platelets with low concentrations of latrunculin A primed platelets for Ca(2+)- or guanosine triphosphate (GTP)-gamma-S-induced alpha-granule secretion. Latrunculin A-dependent alpha-granule secretion was inhibited by antibodies directed at vesicle-associated membrane protein (VAMP), demonstrating that latrunculin A supports soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) protein-dependent membrane fusion. These results indicate that the actin cytoskeleton interferes with platelet exocytosis and differentially regulates alpha-granule and dense granule secretion.
Subject(s)
Actins/metabolism , Blood Platelets/cytology , Blood Platelets/metabolism , Cell Degranulation , Cytoplasmic Granules/metabolism , Cytoskeleton/physiology , Actins/ultrastructure , Blood Platelets/drug effects , Blood Platelets/ultrastructure , Bridged Bicyclo Compounds, Heterocyclic/agonists , Cell Degranulation/drug effects , Cytochalasins/pharmacology , Cytoplasmic Granules/drug effects , Cytoplasmic Granules/ultrastructure , Cytoskeleton/drug effects , Cytoskeleton/ultrastructure , Humans , Kinetics , Microscopy, Electron, Transmission , Peptide Fragments/pharmacology , SNARE Proteins , Tetradecanoylphorbol Acetate/pharmacology , Thiazoles/agonists , Thiazolidines , Vesicular Transport Proteins/metabolismSubject(s)
Adenocarcinoma/pathology , Kidney Neoplasms/pathology , Adenocarcinoma/complications , Adenocarcinoma/surgery , Adenocarcinoma/ultrastructure , Adult , Female , Humans , Hypertension, Renal/etiology , Juxtaglomerular Apparatus , Kidney Neoplasms/complications , Kidney Neoplasms/surgery , Kidney Neoplasms/ultrastructure , NephrectomyABSTRACT
Mouse embryos genetically null for the alphav integrin subunit develop intracerebral hemorrhages at midgestation and die shortly after birth. A key question is whether the hemorrhage arises from primary defects in vascular endothelial cells or pericytes or from other causes. We have previously reported normal initiation of cerebral vessels comprising branched tubes of endothelial cells. Here we show that the onset of hemorrhage is not due to defects in pericyte recruitment. Additionally, most alphav-null vessels display ultrastructurally normal endothelium-pericyte associations and normal interendothelial cell junctions. Thus, endothelial cells and pericytes appear to establish their normal relationships in cerebral microvessels. However, by both light and electron microscopy, we detected defective associations between cerebral microvessels and the surrounding brain parenchyma, composed of neuroepithelial cells, glia, and neuronal precursors. These data suggest a novel role for alphav integrins in the association between cerebral microvessels and central nervous system parenchymal cells.