The major objective of these studies is to increase our understanding of how epidermal cells interact with the extracellular matrix (ECM) during wound closure. Thus, the behavior of adult newt (Notophthalmus viridescens) epidermal cells in wounded limbs will be analyzed as the cells migrate over plastic or nitrocellulose implants coated with collagen, laminin, fibrinogen (FGN), or fibronectin (FN). To learn more about the molecular features that epidermal cells recognize in FGN and FN, proteolytic fragments of each molecule will be tested as migration substrates and/or soluble inhibitors of migration over the intact molecule. In the case of FN, monoclonal antibodies will also be tested as inhibitors of migration over the intact molecule to determine if epidermal cells utilize the fibroblast binding site of FN. To begin characterizing the cell surface receptors which might be involved in recognition of ECM molecules, binding studies will be conducted using latex beads coated with various ECM proteins to determine if the receptors on the upper surface of migrating cells vary in relation to the substrate under the cells. Lectins and glycosidases will be tested as migration substrates to determine if bonds between cell surface saccharides and the substrate could be involved in migration. In addition, adhesion assays will be conducted using dissociated cells in an attempt to demonstrate interaction of epidermal cells with migration-promoting ECM proteins in a more defined environment than a wound. To analyze certain consequences of substrate recognition, immunohistochemical techniques will be used to study the expression of basement membrane zone proteins as epidermal cells migrate over implants coated with various ECM molecules. Finally, experiments will be conducted to determine if epidermal cells lose their dependence on the pre-existing ECM during migration.

Agency
National Institute of Health (NIH)
Institute
National Institute of Arthritis and Musculoskeletal and Skin Diseases (NIAMS)
Type
Research Project (R01)
Project #
5R01AR027940-08
Application #
3155593
Study Section
General Medicine A Subcommittee 2 (GMA)
Project Start
1982-04-01
Project End
1990-03-31
Budget Start
1989-04-01
Budget End
1990-03-31
Support Year
8
Fiscal Year
1989
Total Cost
Indirect Cost
Name
University of Tennessee Health Science Center
Department
Type
Schools of Medicine
DUNS #
941884009
City
Memphis
State
TN
Country
United States
Zip Code
38163
Donaldson, D J; Mahan, J T; Yang, H et al. (1995) Integrin and phosphotyrosine expression in normal and migrating newt keratinocytes. Anat Rec 241:49-58
Donaldson, D J; Mahan, J T; Tsilibary, E C et al. (1994) Migratory interaction of amphibian epidermal cells with components of the basement membrane. J Cell Physiol 158:79-86
Donaldson, D J; Mahan, J T; Amrani, D L et al. (1994) Further studies on the interaction of migrating keratinocytes with fibrinogen. Cell Adhes Commun 2:299-308
Mahan, J T; Donaldson, D J (1992) Divalent cations and extracellular matrix receptor function during newt epidermal cell migration. J Cell Sci 101 ( Pt 1):173-81
Donaldson, D J; Mahan, J T; Yang, H et al. (1991) Tenascin localization in skin wounds of the adult newt Notophthalmus viridescens. Anat Rec 230:451-9
Donaldson, D J; Mahan, J T; Amrani, D et al. (1989) Fibrinogen-mediated epidermal cell migration: structural correlates for fibrinogen function. J Cell Sci 94 ( Pt 1):101-8
Donaldson, D J; Mahan, J T; Smith Jr, G N (1988) Newt epidermal cell migration over collagen and fibronectin involves different mechanisms. J Cell Sci 90 ( Pt 2):325-33
Mahan, J T; Donaldson, D J (1988) A serum-free primary culture system for studying cell-substrate interactions during newt epidermal cell migration. In Vitro Cell Dev Biol 24:1023-30
Atnip, K D; Hade, E P; Donaldson, D J (1988) N-acetylglucosamine binding activity in extracts of adult newt skin. Comp Biochem Physiol A Comp Physiol 90:475-9
Atnip, K D; Mahan, J T; Donaldson, D J (1987) Role of carbohydrates in cell-substrate interactions during newt epidermal cell migration. J Exp Zool 243:461-71

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