| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
|
|
|
|||||||
|
|||||||||
1 From the University of Texas Southwestern Medical Center at Dallas; and the 2 University of Cincinnati, Ohio.
Abstract
PURPOSE. Recent studies indicate that transforming growth factor
(TGF)ß is a potent inducer of corneal myofibroblast
differentiation and expression of smooth muscle–specific, 
-actin
(-SMA). Although TGFß is known to enhance synthesis of
extracellular matrix proteins and receptors, little is known about how
it modulates the expression of smooth muscle proteins in nonmuscle
cells. The purpose of this study was to identify the role of
Arg-Gly-Asp (RGD)-dependent tyrosine phosphorylation in regulating
-SMA gene expression and ultimately myofibroblast development.
METHODS. Because cell culture in serum-containing media mimics myofibroblast
transformation, all experiments were performed on freshly isolated
rabbit keratocytes plated in defined, serum-free media. Cells were
exposed to TGFß (1 ng/ml), Gly-Arg-Gly-Asp-D-Ser-Pro
(GRGDdSP, 50 µM), Gly-Arg-AL-Asp-Ser-Pro (GRADSP; 100 µM), or
herbimycin A (0.1–10 nM) at 24 hours (sparse) or 7 days (confluent).
Cells were evaluated by immunocytochemistry and proteins and RNA
collected for western and northern blot analyses using antibodies
specific for -SMA, fibronectin, focal adhesion proteins, and
phosphotyrosine (clones 4G10 and PY20); and probes directed against
rabbit -SMA. All experiments were repeated at least three times.
RESULTS. Keratocytes exposed to TGFß showed expression of -SMA
that coincided with the intracellular reorganization of the actin
cytoskeleton and the extracellular assembly of fibronectin fibrils.
Addition of RGD containing but not control peptides blocked the
organization of intracellular actin, extracellular fibronectin, and
-SMA protein and mRNA. Immunoprecipitation of cell proteins with
4G10 or PY20 identified the TGFß-associated tyrosine
phosphorylation of paxillin, pp125fak, p130, PLC
, and
tensin, which was blocked by addition of GRGDdSP. Addition of
herbimycin A to keratocytes exposed to TGFß showed a
dose-dependent loss of -SMA protein and mRNA which correlated with
loss of tyrosine phosphorylation, absence of actin reorganization, and
fibronectin assembly.
CONCLUSIONS. The data suggest that TGFß-mediated -SMA gene
expression leading to myofibroblast transformation may involve an
RGD-dependent phosphotyrosine signal transduction
pathway.
This article has been cited by other articles:
|
|
 |
|
  L. J. Dawes, J. A. Eldred, I. K. Anderson, M. Sleeman, J. R. Reddan, G. Duncan, and I. M. Wormstone TGF{beta}-Induced Contraction Is Not Promoted by Fibronectin-Fibronectin Receptor Interaction, or {alpha}SMA Expression Invest. Ophthalmol. Vis. Sci., February 1, 2008; 49(2): 650 - 661. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. M. Bernstein, S. S. Twining, D. J. Warejcka, E. Tall, and S. K. Masur Urokinase Receptor Cleavage: A Crucial Step in Fibroblast-to-Myofibroblast Differentiation Mol. Biol. Cell, July 1, 2007; 18(7): 2716 - 2727. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. Meltendorf, G. J. Burbach, J. Buhren, R. Bug, C. Ohrloff, and T. Deller Corneal Femtosecond Laser Keratotomy Results in Isolated Stromal Injury and Favorable Wound-Healing Response Invest. Ophthalmol. Vis. Sci., May 1, 2007; 48(5): 2068 - 2075. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
E. Guerriero, J. Chen, Y. Sado, R. R. Mohan, S. E. Wilson, J. L. Funderburgh, and N. SundarRaj Loss of Alpha3(IV) Collagen Expression Associated with Corneal Keratocyte Activation Invest. Ophthalmol. Vis. Sci., February 1, 2007; 48(2): 627 - 635. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 G Perrella, P Brusini, R Spelat, P Hossain, A Hopkinson, and H S Dua Expression of haematopoietic stem cell markers, CD133 and CD34 on human corneal keratocytes Br. J. Ophthalmol., January 1, 2007; 91(1): 94 - 99. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 R. S. Greenberg, A. M. Bernstein, M. Benezra, I. H. Gelman, L. Taliana, and S. K. Masur FAK-dependent regulation of myofibroblast differentiation FASEB J, May 1, 2006; 20(7): 1006 - 1008. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. Izumi, D. Kurosaka, T. Iwata, Y. Oguchi, Y. Tanaka, Y. Mashima, and K. Tsubota Involvement of Insulin-like Growth Factor-I and Insulin-like Growth Factor Binding Protein-3 in Corneal Fibroblasts during Corneal Wound Healing Invest. Ophthalmol. Vis. Sci., February 1, 2006; 47(2): 591 - 598. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Y. Kim, M. J. Kim, T.-i. Kim, H.-j. Choi, J. H. Pak, and H. Tchah A Femtosecond Laser Creates a Stronger Flap than a Mechanical Microkeratome Invest. Ophthalmol. Vis. Sci., February 1, 2006; 47(2): 599 - 604. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Yoshida, S. Shimmura, J. Shimazaki, N. Shinozaki, and K. Tsubota Serum-Free Spheroid Culture of Mouse Corneal Keratocytes Invest. Ophthalmol. Vis. Sci., May 1, 2005; 46(5): 1653 - 1658. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P. Ottino, J. He, T. W. Axelrad, and H. E. P. Bazan PAF-Induced Furin and MT1-MMP Expression Is Independent of MMP-2 Activation in Corneal Myofibroblasts Invest. Ophthalmol. Vis. Sci., February 1, 2005; 46(2): 487 - 496. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
L. Taliana, M. Benezra, R. S. Greenberg, S. K. Masur, and A. M. Bernstein ZO-1: Lamellipodial Localization in a Corneal Fibroblast Wound Model Invest. Ophthalmol. Vis. Sci., January 1, 2005; 46(1): 96 - 103. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. E. Wilson, R. R. Mohan, M. Netto, V. Perez, D. Possin, J. Huang, R. Kwon, A. Alekseev, and J. P. Rodriguez-Perez RANK, RANKL, OPG, and M-CSF Expression in Stromal Cells during Corneal Wound Healing Invest. Ophthalmol. Vis. Sci., July 1, 2004; 45(7): 2201 - 2211. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 R. B. Hough and J. Piatigorsky Preferential Transcription of Rabbit Aldh1a1 in the Cornea: Implication of Hypoxia-Related Pathways Mol. Cell. Biol., February 1, 2004; 24(3): 1324 - 1340. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. G. Ryan, L. Taliana, L. Sun, Z.-G. Wei, S. K. Masur, and R. M. Lavker Involvement of S100A4 in Stromal Fibroblasts of the Regenerating Cornea Invest. Ophthalmol. Vis. Sci., October 1, 2003; 44(10): 4255 - 4262. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. C. Hubchak, C. E. Runyan, J. I. Kreisberg, and H. W. Schnaper Cytoskeletal Rearrangement and Signal Transduction in TGF-{beta}1-Stimulated Mesangial Cell Collagen Accumulation J. Am. Soc. Nephrol., August 1, 2003; 14(8): 1969 - 1980. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. V. Jester, J. Huang, S. Fisher, J. Spiekerman, J. H. Chang, W. E. Wright, and J. W. Shay Myofibroblast Differentiation of Normal Human Keratocytes and hTERT, Extended-Life Human Corneal Fibroblasts Invest. Ophthalmol. Vis. Sci., May 1, 2003; 44(5): 1850 - 1858. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Saika, T. Miyamoto, S.-i. Tanaka, T. Tanaka, I. Ishida, Y. Ohnishi, A. Ooshima, T. Ishiwata, G. Asano, T.-i. Chikama, et al. Response of Lens Epithelial Cells to Injury: Role of Lumican in Epithelial-Mesenchymal Transition Invest. Ophthalmol. Vis. Sci., May 1, 2003; 44(5): 2094 - 2102. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
B. L. Berryhill, R. Kader, B. Kane, D. E. Birk, J. Feng, and J. R. Hassell Partial Restoration of the Keratocyte Phenotype to Bovine Keratocytes Made Fibroblastic by Serum Invest. Ophthalmol. Vis. Sci., November 1, 2002; 43(11): 3416 - 3421. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J.-T. Kim and C.-K. Joo Involvement of Cell-Cell Interactions in the Rapid Stimulation of Cas Tyrosine Phosphorylation and Src Kinase Activity by Transforming Growth Factor-beta 1 J. Biol. Chem., August 23, 2002; 277(35): 31938 - 31948. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 L. Ronnov-Jessen, R. Villadsen, J. C. Edwards, and O. W. Petersen Differential Expression of a Chloride Intracellular Channel Gene, CLIC4, in Transforming Growth Factor-{beta}1-Mediated Conversion of Fibroblasts to Myofibroblasts Am. J. Pathol., August 1, 2002; 161(2): 471 - 480. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
L. You and F. E. Kruse Differential Effect of Activin A and BMP-7 on Myofibroblast Differentiation and the Role of the Smad Signaling Pathway Invest. Ophthalmol. Vis. Sci., January 1, 2002; 43(1): 72 - 81. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. L. Funderburgh, M. L. Funderburgh, M. M. Mann, L. Corpuz, and M. R. Roth Proteoglycan Expression during Transforming Growth Factor beta -induced Keratocyte-Myofibroblast Transdifferentiation J. Biol. Chem., November 16, 2001; 276(47): 44173 - 44178. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
O. Maltseva, P. Folger, D. Zekaria, S. Petridou, and S. K. Masur Fibroblast Growth Factor Reversal of the Corneal Myofibroblast Phenotype Invest. Ophthalmol. Vis. Sci., October 1, 2001; 42(11): 2490 - 2495. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P. A. Folger, D. Zekaria, G. Grotendorst, and S. K. Masur Transforming Growth Factor-{beta}-Stimulated Connective Tissue Growth Factor Expression during Corneal Myofibroblast Differentiation Invest. Ophthalmol. Vis. Sci., October 1, 2001; 42(11): 2534 - 2541. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. E. Wilson, R. R. Mohan, J.-W. Hong, J.-S. Lee, R. Choi, and R. R. Mohan The Wound Healing Response After Laser In Situ Keratomileusis and Photorefractive Keratectomy: Elusive Control of Biological Variability and Effect on Custom Laser Vision Correction Arch Ophthalmol, June 1, 2001; 119(6): 889 - 896. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. Nagano, J.-L. Hao, M. Nakamura, N. Kumagai, M. Abe, T. Nakazawa, and T. Nishida Stimulatory Effect of Pseudomonal Elastase on Collagen Degradation by Cultured Keratocytes Invest. Ophthalmol. Vis. Sci., May 1, 2001; 42(6): 1247 - 1253. [Abstract] [Full Text]
|
|
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
