Protein Kinase C Activation Stimulates Mesenchymal Stem Cell Adhesion through Activation of Focal Adhesion Kinase

Background Tyrosine protein kinase proteins exert a prominent control on signaling pathways and may couple rapid events, such as action potential and neurotransmitter release, to long-lasting changes in synaptic strength and survival. Whether anesthetics modulate tyrosine kinase activity remains unknown. The aim of the current study was therefore to examine the effects of intravenous and volatile anesthetics on the phosphorylation of focal adhesion kinase (ppFAK), a functionally important nonreceptor tyrosine kinase, in the rat hippocampus. Methods Phosphorylation of ppFAK was examined in hippocampal slices by immunoblotting with both antiphosphotyrosine and specific anti-ppFAK antibodies. Experiments were performed in the absence (control) or presence of various concentrations of pharmacologic or anesthetic agents or both. Results Clinically relevant concentrations of thiopental, propofol, etomidate, isoflurane, sevoflurane, and desflurane induced a concentration-related increase in tyrosine phosphorylation. In contrast, ketamine (up to 100 microm) and the nonimmobilizer F6 (1,2-dichlorohexafluorocyclobutane, 25 microm) did not significantly affect ppFAK phosphorylation. The anesthetic-induced increase in ppFAK phosphorylation was blocked by GF 109203X, RO 318220, and chelerythrin (100 microm), three structurally distinct inhibitors of protein kinase C and U 73122 (50 microm), an inhibitor of phospholipase C. The propofol- and isoflurane-induced increase in ppFAK phosphorylation was reversible and showed nonadditivity of effects with phorbol 12-myristate 13-acetate (an activator of protein kinase C, 0.1 microm). In contrast, ketamine (up to 100 microm), MK801 (10 microm, an N-methyl-d-aspartate receptor antagonist), bicuculline (10 microm, a gamma-aminobutyric acid type A receptor antagonist), and dantrolene (30 microm, an inhibitor of the ryanodine receptor) were ineffective in blocking anesthetic-induced activation of tyrosine phosphorylation. Conclusion Except for ketamine, anesthetic agents markedly increase tyrosine phosphorylation of ppFAK in the rat hippocampus, most likely via the phospholipase C-protein kinase C pathway, whereas the nonimmobilizer F6 does not. These results suggest that ppFAK represents a target for anesthetic action in the brain.

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Endothelin-1 Stimulates Tyrosine Phosphorylation of p125 Focal Adhesion Kinase via Protein Kinase C in Neonatal Cardiac Myocytes

The Japanese Journal of Pharmacology ◽

10.1016/s0021-5198(19)41198-0 ◽

1998 ◽

Vol 76 ◽

pp. 272

Author(s):

Rikako Yamauchi ◽

Tomoko Hoshino ◽

Kohei Kikkawa ◽

Hideo Yabana ◽

Sakae Murata

Keyword(s):

Protein Kinase C ◽

Protein Kinase ◽

Focal Adhesion Kinase ◽

Tyrosine Phosphorylation ◽

Cardiac Myocytes ◽

Focal Adhesion ◽

Endothelin 1 ◽

Kinase C

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Protein kinase C regulates alpha v beta 5-dependent cytoskeletal associations and focal adhesion kinase phosphorylation.

The Journal of Cell Biology ◽

10.1083/jcb.134.5.1323 ◽

1996 ◽

Vol 134 (5) ◽

pp. 1323-1332 ◽

Cited By ~ 130

Author(s):

J M Lewis ◽

D A Cheresh ◽

M A Schwartz

Keyword(s):

Protein Kinase C ◽

Protein Kinase ◽

Focal Adhesion Kinase ◽

Focal Adhesion ◽

Cytoskeletal Proteins ◽

Kinase C ◽

Focal Adhesion Proteins ◽

Mediate Cell ◽

Alpha V Beta 3 ◽

Kinase Phosphorylation

Integrins alpha v beta 3 and alpha v beta 5 both mediate cell adhesion to vitronectin yet trigger different postligand binding events. Integrin alpha v beta 3 is able to induce cell spreading, migration, angiogenesis, and tumor metastasis without additional stimulators, whereas alpha v beta 5 requires exogenous activation of protein kinase C (PKC) to mediate these processes. To investigate this difference, the ability of beta 3 or beta 5 to induce colocalization of intracellular proteins was assessed by immunofluorescence in hamster CS-1 melanoma cells. We found that alpha v beta 5 induced colocalization of talin, alpha-actinin, tensin, and actin very weakly relative to alpha v beta 3. alpha v beta 5 was able to efficiently induce colocalization of focal adhesion kinase (FAK); however, it was unable to increase phosphorylation of FAK on tyrosine. Activation of PKC by adding phorbol ester to alpha v beta 5-expressing cells induced spreading, increased colocalization of alpha-actinin, tensin, vinculin, p130cas and actin, and triggered tyrosine phosphorylation of FAK. Unexpectedly, talin colocalization remained low even after activation of PKC. Treatment of cells with the PKC inhibitor calphostin C inhibited spreading and the colocalization of talin, alpha-actinin, tensin, and actin for both alpha v beta 3 and alpha v beta 5. We conclude that PKC regulates localization of cytoskeletal proteins and phosphorylation of FAK induced by alpha v beta 5. Our results also show that FAK can be localized independent of its phosphorylation and that cells can spread and induce localization of other focal adhesion proteins in the absence of detectable talin.

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Bombesin stimulation of p125 focal adhesion kinase tyrosine phosphorylation. Role of protein kinase C, Ca2+ mobilization, and the actin cytoskeleton

Journal of Biological Chemistry ◽

10.1016/s0021-9258(19)85236-5 ◽

1993 ◽

Vol 268 (19) ◽

pp. 14261-14268

Author(s):

J. Sinnett-Smith ◽

I. Zachary ◽

A.M. Valverde ◽

E. Rozengurt

Keyword(s):

Protein Kinase C ◽

Protein Kinase ◽

Actin Cytoskeleton ◽

Focal Adhesion Kinase ◽

Tyrosine Phosphorylation ◽

Focal Adhesion ◽

Kinase C ◽

Stimulation Of

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Possible role of protein kinase C in the regulation of intracellular stability of focal adhesion kinase in mouse 3T3 cells

FEBS Letters ◽

10.1016/0014-5793(95)01014-6 ◽

1995 ◽

Vol 373 (2) ◽

pp. 135-140 ◽

Cited By ~ 12

Author(s):

Akira Mogi ◽

Mika Hatai ◽

Hisae Soga ◽

Seiichi Takenoshita ◽

Yukio Nagamachi ◽

...

Keyword(s):

Protein Kinase C ◽

Protein Kinase ◽

Focal Adhesion Kinase ◽

Focal Adhesion ◽

3T3 Cells ◽

Kinase C

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Protein kinase C dependent induction of pp125FAK in monocytes by colony stimulating factor-GM: evidence for a synergistic effect by the cytokine and 1,25 dihydroxyvitamin D3

Journal of Endocrinology ◽

10.1677/joe.0.152r019 ◽

1997 ◽

Vol 152 (2) ◽

pp. R19-R22 ◽

Cited By ~ 1

Author(s):

Raed M Kanan ◽

Hersha Rathod ◽

Harish K Datta

Keyword(s):

Protein Kinase C ◽

Protein Kinase ◽

Focal Adhesion Kinase ◽

Focal Adhesion ◽

Prolonged Exposure ◽

Dihydroxyvitamin D3 ◽

Dihydroxyvitamin D ◽

Kinase C ◽

Gf 109203X

Abstract The differentiation of monocytes into osteoclasts has been recently achieved in vitro in a suitable milieu containing morphogens that includes 1,25 dihydroxyvitamin D3, colony stimulating factors, interleukins and the presence of cells of osteoblastic lineage. However, the precise role of these factors in the osteoclastic differentiation process has not yet been examined. Since our previous studies have shown that osteoclasts express a much higher level of focal adhesion kinase (pp125FAK) than cells of macrophage/monocytic lineage, the present study was carried out to ascertain which morphogens are involved in increasing the expression of the kinase during the differentiation of monocytes to osteoclasts. We demonstrate that a marked increase in the expression of pp125FAK occurs only after prolonged exposure to hCSF-GM and combination of hCSF-GM and 1,25 (OH)2 D3. The hCSF-GM was found to be a more potent stimulator of pp125FAK induction than 1,25 (OH)2 D3; interestingly, the presence of both hCSF-GM and 1,25 (OH)2 D3 showed co-operative effect. Furthermore, the presence of a protein kinase C inhibitor, bisindolylmaleimide (GF 109203X), blocked hCSF-GM-mediated induction of focal adhesion kinase, implicating an important role for protein kinase C in the induction of pp125FAK.

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