Association of Focal Adhesion Kinase with Fibronectin and Paxillin Is Required for Precartilage Condensation of Chick Mesenchymal Cells

Bang, Ok‐Sun; Kim, Eun-Jung; Chung, Jae Gwon; Lee, Sun-Ryung; Park, Tae Kyu; Kang, Shin‐Sung

doi:10.1006/bbrc.2000.3831

Cited by 60 publications

(38 citation statements)

References 33 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Phosphatases that affect chondrogenesis [51][52][53] and mineralization [54] in similar cultures also have been reported, and it is likely that at the early stages of differentiation the effects are on intracellular events as most of the phosphatases signal across the membrane. There may also be effects of blocking CK2 on such events.…”

Section: Discussionmentioning

confidence: 93%

Modulation of extracellular matrix protein phosphorylation alters mineralization in differentiating chick limb-bud mesenchymal cell micromass cultures

Boskey

Doty

Kudryashov

et al. 2008

Bone

View full text Add to dashboard Cite

Protein phosphorylation and dephosphorylation are important regulators of cellular and extracellular events. The purpose of this study was to define how these events regulate cartilage matrix calcification in a cell culture system that mimics endochondral ossification. The presence of casein kinase II (CK2), an enzyme known to phosphorylate matrix proteins, was confirmed by immunohistochemistry. The importance of phosphoprotein phosphorylation and dephosphorylation was examined by comparing effects of inhibiting CK2 or phosphoprotein phosphatases on mineral accretion relative to untreated mineralizing controls. Specific inhibitors were added to differentiating chick limb bud mesenchymal cell micromass cultures during the development of a mineralized matrix at the times of cell differentiation, proliferation, formation of the mineralized matrix, or proliferation of the mineral crystals. The mineralizing media for these cultures contained 4mM inorganic phosphate and no organic-phosphate esters; control cultures had 1mM inorganic phosphate. Mineralization was monitored based on 45 Ca uptake and infrared characterization of the mineral; cell viability was assessed by three independent methods. Treatments that caused cell toxicity were excluded from the analysis.Inhibition of CK2 activity with apigenin or CK2 inhibitor II reduced the rate of mineral deposition, but did not block mineral accretion. Effects were greatest during the time of mineralized matrix formation. Inhibition of phosphoprotein phosphatase activities with okadaic acid, calyculin A, and microcystin LR, at early time points also markedly inhibited mineral accretion. Inhibition after mineralization had commenced increased the mineral yield. Levamisole, an alkaline phosphatase inhibitor, had no effect on mineral accretion in this system, suggesting the involvement of other phosphatases. Adding additional inorganic phosphate to the inhibited cultures after mineralization had started, but not earlier, reversed the inhibition indicating the phosphatases were, in part, providing a source of inorganic phosphate.To characterize the roles of specific phosphoproteins blocking studies were performed. Blocking with anti-osteopontin antibody confirmed osteopontin's previously reported role as a mineralization inhibitor. Blocking antibodies to bone sialoprotein added from day 9 or on days 9 and 11 retarded mineralization, supporting its role as a mineralization nucleator. Antibodies to osteonectin slightly stimulated early mineralization, but had no effect after the time that initial mineral deposition occurs. Taken together, the results of this study demonstrate the importance of the phosphorylation state of extracellular matrix proteins in regulating mineralization in this culture system.

show abstract

Section: Discussionmentioning

confidence: 93%

Modulation of extracellular matrix protein phosphorylation alters mineralization in differentiating chick limb-bud mesenchymal cell micromass cultures

Boskey

Doty

Kudryashov

et al. 2008

Bone

View full text Add to dashboard Cite

show abstract

“…Numerous signaling pathways that lie downstream of integrins or are activated by chondrogenic factors have been linked to various stages of this process. Activation of focal adhesion kinase (FAK) and paxillin occurs during pre-cartilage condensation (Bang et al, 2000). Extracellular-signal-regulated kinases 1 and 2 (ERKs1/2, also known as MAPK3 and MAPK1, Fibronectin and chondrogenesis 3707 respectively) are activated by binding of preadipocyte factor 1 (Pref-1, also known as DLK1, a transmembrane protein that contains EGF-like repeats) to fibronectin, which leads to upregulation of SOX9 and inhibition of adipogenesis .…”

Section: Ecm Deposition During Chondrogenesismentioning

confidence: 99%

Fibronectin and stem cell differentiation – lessons from chondrogenesis

Singh

Schwarzbauer

2012

Journal of Cell Science

168

156

View full text Add to dashboard Cite

SummaryThe extracellular matrix (ECM) is an intricate network of proteins that surrounds cells and has a central role in establishing an environment that is conducive to tissue-specific cell functions. In the case of stem cells, this environment is the stem cell niche, where ECM signals participate in cell fate decisions. In this Commentary, we describe how changes in ECM composition and mechanical properties can affect cell shape and stem cell differentiation. Using chondrogenic differentiation as a model, we examine the changes in the ECM that occur before and during mesenchymal stem cell differentiation. In particular, we focus on the main ECM protein fibronectin, its temporal expression pattern during chondrogenic differentiation, its potential effects on functions of differentiating chondrocytes, and how its interactions with other ECM components might affect cartilage development. Finally, we discuss data that support the possibility that the fibronectin matrix has an instructive role in directing cells through the condensation, proliferation and/or differentiation stages of cartilage formation.

show abstract

“…Outside-in signaling through integrins can result in Src dephosphorylation of its inhibitory Y527 residue, which in turn increases its activation. 27,28 The critical role of detachment from a substratum for the process of motility has not been studied as extensively as has the process of adherence. During migration of epithelial cells across an extracellular matrix, levels of membrane-associated FAK and its tyrosine phosphorylation are decreased.…”

mentioning

confidence: 99%

Protein phosphatase‐2A regulates endothelial cell motility and both the phosphorylation and the stability of focal adhesion complexes

Young

Kolesiak

Meisinger

2002

Intl Journal of Cancer

View full text Add to dashboard Cite

Association of Focal Adhesion Kinase with Fibronectin and Paxillin Is Required for Precartilage Condensation of Chick Mesenchymal Cells

Cited by 60 publications

References 33 publications

Modulation of extracellular matrix protein phosphorylation alters mineralization in differentiating chick limb-bud mesenchymal cell micromass cultures

Modulation of extracellular matrix protein phosphorylation alters mineralization in differentiating chick limb-bud mesenchymal cell micromass cultures

Fibronectin and stem cell differentiation – lessons from chondrogenesis

Protein phosphatase‐2A regulates endothelial cell motility and both the phosphorylation and the stability of focal adhesion complexes

Contact Info

Product

Resources

About