The goal of the proposal is to determine the role of fibronectin in the differentiation of chondrocytes. Since differentiation of mesenchymal cells into chondrocytes is one of the earliest steps in endochondral bone formation and bone fracture repair, the studies proposed should lead to an increased understanding of chondrogenic expression and bone morphogenesis. It has been demonstrated that cellular fibronectin is one of the chief determinants of chondrogenic expression. In the presence of fibronectin, chondroblasts remain undifferentiated; while the removal of fibronectin or the addition of antibodies of fibronectin results in chondrogenic expression. Based on our current understanding of the molecular biology of fibronectin, we propose to examine the molecular mechanism involved in the control of chondrogenic expression by fibronectin. We will determine the region of fibronectin responsible for its effects on the differentiation of chondrocytes by the isolation of appropriate proteolytic fragments of fibronectin. The fibronectin fragment isolated will be analyzed in order to determine the identity of the receptor responsible for chondrogenic expression. The cell surface molecular responsible for the differentiative action of fibronectin will be isolated. Intrinsic membrane molecules which participate in fibronectin-mediated cell adhesion will be examined by biochemical and biophysical means.

Agency
National Institute of Health (NIH)
Institute
National Institute of Dental & Craniofacial Research (NIDCR)
Type
Research Project (R01)
Project #
5R01DE008144-04
Application #
3221914
Study Section
Oral Biology and Medicine Subcommittee 1 (OBM)
Project Start
1988-07-01
Project End
1993-06-30
Budget Start
1991-07-01
Budget End
1993-06-30
Support Year
4
Fiscal Year
1991
Total Cost
Indirect Cost
Name
University of Texas Health Science Center San Antonio
Department
Type
Schools of Medicine
DUNS #
800772162
City
San Antonio
State
TX
Country
United States
Zip Code
78229
Giambernardi, T A; Sakaguchi, A Y; Gluhak, J et al. (2001) Neutrophil collagenase (MMP-8) is expressed during early development in neural crest cells as well as in adult melanoma cells. Matrix Biol 20:577-87
Giambernardi, T A; Klebe, R J (2000) Relative reverse transcription-polymerase chain reaction. Methods Mol Biol 137:51-8
Klebe, R J; Rodriguez, S A; VerBeek, M L et al. (1999) Simple method for ""hot-starting"" RT-PCR. Biotechniques 27:1108-10
Grant, G M; Giambernardi, T A; Grant, A M et al. (1999) Overview of expression of matrix metalloproteinases (MMP-17, MMP-18, and MMP-20) in cultured human cells. Matrix Biol 18:145-8
Taylor, G P; Troyer, D A; Giambernardi, T A et al. (1998) Extraction of RNA from single frozen sections. J Pathol 184:332-5
Giambernardi, T A; Grant, G M; Taylor, G P et al. (1998) Overview of matrix metalloproteinase expression in cultured human cells. Matrix Biol 16:483-96
Giambernardi, T A; Rodeck, U; Klebe, R J (1998) Bovine serum albumin reverses inhibition of RT-PCR by melanin. Biotechniques 25:564-6
Giambernardi, T A; Klebe, R J (1997) Use of oxygen-permeable silicone rubber pouches for growing mass cultures of bacteria. Lett Appl Microbiol 24:207-10
Garcia, M A; Klebe, R J (1997) Affinity chromatography of RNase inhibitor. Mol Biol Rep 24:231-3
Klebe, R J; Grant, G M; Grant, A M et al. (1996) RT-PCR without RNA isolation. Biotechniques 21:1094-100

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