1.CD98 activation increases surface expression and clusteringof beta 1 integrins in MCF-7 cells through FAK/Src- and cytoskeleton-independent mechanisms.
Experimental & Molecular Medicine 2008;40(3):261-270
CD98, a disulfide-linked 125-kDa heterodimeric type II transmembrane glycoprotein, regulates beta 1 integrin- mediated cell adhesion. However, the molecular mechanisms underlying CD98-mediated activation of beta 1 integrin are presently unclear. In this study, the effects of CD98 signaling on the expression and clustering of beta 1 integrin were investigated. Activation of CD98 augmented surface expression of beta 1 integrin on MCF-7 cells. Cross-linking CD98 induced clustering of beta 1 integrins. Inhibition of phosphorylation of focal adhesion kimase (FAK) by PP2, an inhibitor of Src family kinase, reduced cell-extracellular matrix adhesion, but not surface expression and clustering of beta1 integrin on MCF-7 cells. This result was confirmed by over-expression of dominant negative forms of FAK. In addition, phalloidin or cytochalasin D inhibited CD98-mediated induction of cell-ECM adhesion, but not surface expression and clustering of b1 integrins. The inhibitory effects of PP2, cytochalasin D or phalloidin on CD98-stimulated cell adhesion were diminished by pretreatment of cells with Mn2+, which is shown to induce conformational change of integrins. These results provide the first evidence that CD98 activation increases not only beta1 integrin affinity but also its surface expression and clustering and the latter is independent of FAK/Src and cytoskeleton.
Antigens, CD29/*biosynthesis/genetics
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Antigens, CD98/agonists/*metabolism
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Cell Line, Tumor
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Cytochalasin D/pharmacology
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Cytoskeleton/drug effects/enzymology
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Focal Adhesion Kinase 2/genetics/*metabolism
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Focal Adhesions/drug effects/enzymology
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Humans
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Microscopy, Confocal
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Multiprotein Complexes/*biosynthesis/genetics
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Mutant Proteins/genetics/metabolism
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Phalloidine/pharmacology
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Phosphorylation/drug effects
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Protein Binding
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Pyrimidines/pharmacology
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Signal Transduction/physiology
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Transfection
2.beta1-integrin-dependent migration of microglia in response to neuron-released alpha-synuclein.
Changyoun KIM ; Eun Deok CHO ; Hyung Koo KIM ; Sungyong YOU ; He Jin LEE ; Daehee HWANG ; Seung Jae LEE
Experimental & Molecular Medicine 2014;46(4):e91-
Chronic neuroinflammation is an integral pathological feature of major neurodegenerative diseases. The recruitment of microglia to affected brain regions and the activation of these cells are the major events leading to disease-associated neuroinflammation. In a previous study, we showed that neuron-released alpha-synuclein can activate microglia through activating the Toll-like receptor 2 (TLR2) pathway, resulting in proinflammatory responses. However, it is not clear whether other signaling pathways are involved in the migration and activation of microglia in response to neuron-released alpha-synuclein. In the current study, we demonstrated that TLR2 activation is not sufficient for all of the changes manifested by microglia in response to neuron-released alpha-synuclein. Specifically, the migration of and morphological changes in microglia, triggered by neuron-released alpha-synuclein, did not require the activation of TLR2, whereas increased proliferation and production of cytokines were strictly under the control of TLR2. Construction of a hypothetical signaling network using computational tools and experimental validation with various peptide inhibitors showed that beta1-integrin was necessary for both the morphological changes and the migration. However, neither proliferation nor cytokine production by microglia was dependent on the activation of beta1-integrin. These results suggest that beta1-integrin signaling is specifically responsible for the recruitment of microglia to the disease-affected brain regions, where neurons most likely release relatively high levels of alpha-synuclein.
Animals
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Antigens, CD29/genetics/*metabolism
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Cell Line, Tumor
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*Cell Movement
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Cells, Cultured
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Culture Media, Conditioned/*pharmacology
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Gene Regulatory Networks
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Humans
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Mice
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Mice, Inbred C57BL
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Microglia/drug effects/metabolism/*physiology
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Neurons/*metabolism
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Rats
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Rats, Sprague-Dawley
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Signal Transduction
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Toll-Like Receptor 2/metabolism
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alpha-Synuclein/*pharmacology