Transforming growth factorβ-mediated corneal myofibroblast differentiation requires actin and fibronectin assembly

被引:0
|
作者
Jester, JV
Huang, JY
Barry-Lane, PA
Kao, WWY
Petroll, WM
Cavanagh, HD
机构
[1] Univ Texas, SW Med Ctr, Dallas, TX 75235 USA
[2] Univ Cincinnati, Cincinnati, OH USA
关键词
D O I
暂无
中图分类号
R77 [眼科学];
学科分类号
100212 ;
摘要
PURPOSE. Recent studies indicate that transforming growth factor (TGF)(beta) is a potent inducer of corneal myofibroblast differentiation and expression of smooth muscle-specific, alpha-actin (alpha-SMA). Although TGF(beta) 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 alpha-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 TGF(beta) (1 ng/ml), Gly-Arg-Gly-Asp-D-Ser-Pro (GRGDdSP, 50 mu M), Gly-Arg-AL-Asp-Ser-Pro (GRADSP; 100 mu M), or herbimycin A (0.1-10 ml) 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 alpha-SMA, fibronectin, focal adhesion proteins, and phosphotyrosine (clones 4G10 and PY20); and probes directed against rabbit alpha-SMA. All experiments were repeated at least three times. RESULTS. Keratocytes exposed to TGF(beta) showed expression of alpha-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 alpha-SMA protein and mRNA. Immunoprecipitation of cell proteins with 4G10 or PY20 identified the TGF(beta)-associated tyrosine phosphorylation of paxillin, pp125(fak), p130, PLC gamma, and tensin, which was blocked by addition of GRGDdSP. Addition of herbimycin A to keratocytes exposed to TGF(beta) showed a dose-dependent loss of alpha-SMA protein and mRNA which correlated with loss of tyrosine phosphorylation, absence of actin reorganization, and fibronectin assembly. CONCLUSIONS. The data suggest that TGF(beta)-mediated alpha-SMA gene expression leading to myofibroblast transformation may involve an RGD-dependent phosphotyrosine signal transduction pathway.
引用
收藏
页码:1959 / 1967
页数:9
相关论文
共 50 条
  • [41] Transforming growth factor-beta 1(TGF-beta 1) induction of myofibroblast functional differentiation.
    Vaughan, MB
    Tomasek, JJ
    FASEB JOURNAL, 1997, 11 (03): : 20 - 20
  • [42] Blue light inhibits transforming growth factor-β1-induced myofibroblast differentiation of human dermal fibroblasts
    Taflinski, Leonie
    Demir, Erhan
    Kauczok, Jens
    Fuchs, Paul Christian
    Born, Matthias
    Suschek, Christoph V.
    Oplaender, Christian
    EXPERIMENTAL DERMATOLOGY, 2014, 23 (04) : 240 - 246
  • [43] Transforming growth factor (TGF)-β1-induced miR-133a inhibits myofibroblast differentiation and pulmonary fibrosis
    Peng Wei
    Yan Xie
    Peter W. Abel
    Yapei Huang
    Qin Ma
    Linghai Li
    Junfeng Hao
    Dennis W. Wolff
    Taotao Wei
    Yaping Tu
    Cell Death & Disease, 10
  • [44] Sequential Analysis of Myofibroblast Differentiation and Transforming Growth Factor-β1/Smad Pathway Activation in Murine Pulmonary Fibrosis
    Usuki, Jiro
    Matsuda, Kuniko
    Azuma, Arata
    Kudoh, Shoji
    Gemma, Akihiko
    JOURNAL OF NIPPON MEDICAL SCHOOL, 2012, 79 (01) : 46 - 59
  • [45] Transforming Growth Factor-Beta1 and Human Gingival Fibroblast-to-Myofibroblast Differentiation: Molecular and Morphological Modifications
    Marconi, Guya D.
    Fonticoli, Luigia
    Rajan, Thangavelu Soundara
    Lanuti, Paola
    Della Rocca, Ylenia
    Pierdomenico, Sante D.
    Trubiani, Oriana
    Pizzicannella, Jacopo
    Diomede, Francesca
    FRONTIERS IN PHYSIOLOGY, 2021, 12
  • [46] Transforming-Growth-Factor-β-Induced Corneal Dystrophies
    Finis, D.
    Stammen, J.
    Geerling, G.
    KLINISCHE MONATSBLATTER FUR AUGENHEILKUNDE, 2014, 231 (06) : 653 - +
  • [47] Suppression of transforming growth factor-beta isoforms, TGF-β receptor type II, and myofibroblast differentiation in cultured human corneal and limbal fibroblasts by amniotic membrane matrix
    Tseng, SCG
    Li, DQ
    Ma, X
    JOURNAL OF CELLULAR PHYSIOLOGY, 1999, 179 (03) : 325 - 335
  • [48] FIBRONECTIN DOES NOT ENHANCE EPIDERMAL GROWTH FACTOR-MEDIATED ACCELERATION OF CORNEAL EPITHELIAL WOUND CLOSURE
    SOONG, HK
    HASSAN, T
    VARANI, J
    HUANG, SCM
    BRENNAN, M
    ARCHIVES OF OPHTHALMOLOGY, 1989, 107 (07) : 1052 - 1054
  • [50] TRANSFORMING GROWTH-FACTOR-BETA STIMULATES COLLAGEN AND FIBRONECTIN SYNTHESIS BY HUMAN CORNEAL STROMAL FIBROBLASTS IN-VITRO
    OHJI, M
    SUNDARRAJ, N
    THOFT, RA
    CURRENT EYE RESEARCH, 1993, 12 (08) : 703 - 709