# CELL-SUBSTRATE INTERACTIONS IN EPIDERMAL CELL MIGRATION

> **NIH NIH R01** · UNIVERSITY OF TENNESSEE HEALTH SCI CTR · 1993 · $154,832

## Abstract

The objective of these studies is to increase our understanding of how
epidermal cells interact with the extracellular matrix (ECM) during wound
closure. The behavior of adult newt (Notophthalmus viridescens) epidermal
cells in wounded limbs will be analyzed as the cells migrate over the
natural wound bed or over implants coated with various ECM proteins,
fragments of ECM proteins, or synthetic peptides modeled after potentially
important regions of ECM proteins. These studies will be supplemented by
adhesion and spreading assays using single cell suspensions of freshly
isolated epidermal cells seeded onto plastic coated with many of the same
substrates as in implant experiments.

More specifically: Novel Fibrinogen (FGN) fragments, cyanogen bromide (CB)
peptides of the alpha chain, and synthetic peptides mimicking the
RGD-containing regions of the alpha chain will be tested as migration
substrates to better localize the migration-supporting active sites in FGN.
The effects of pretreating FGN and FGN fragment-coated implants with
monoclonal antibodies against known regions of the alpha chain and against
the platelet binding site in the gamma chain will be determined. The
ability of the previously mentioned synthetic peptides to act as
competitive inhibitors of migration on FGN and FGN fragments will also be
assessed. To better localize the active site(s) controlling adhesion,
spreading, and migration on fibronectin (FN), recombinant DNA-generated FN
fragments with known amino acid deletions in domains III and IV will be
tested as migration substrates. Adhesion and spreading assays will be
performed on these same fragments as well as on a heparin-binding fragment
that contains most of the III CS region. To localize epidermal recognition
site(s) in type IV collagen, adhesion, spreading, and migration assays will
be conducted using as substrates: isolated type IV structural domains,
isolated alpha chains, pepsin and CB-generated alpha chain fragments, and
synthetic peptides modeled after certain regions of the alpha chains. To
determine the epidermal interactive potential of laminin, entactin, and
heparan sulfate proteoglycan, each protein will be tested as a substrate
in adhesion, spreading, and migration assays. Freshly wounded limbs will
be treated with polyclonal antibodies to collagen, FGN and FN to determine
the relative involvement of each protein in wound closure in vivo. To
characterize the ECM receptors on epidermal cells, polyclonal antibodies
against the beta subunit of known integrins will be used for
immunoprecipitation and immunoblot analysis of epidermal cell extracts and
to inhibit adhesion, spreading, and migration on various ECM proteins.
Migration-inhibiting polyclonal antibodies against newt epidermal cell
membranes will be used to identify cell surface proteins involved in ECM
interaction. FN receptors will be purified from epidermal extracts by
affinity chromatography on the alpha chymotrypsin 120 kD cell-binding
fragment of F...

## Key facts

- **NIH application ID:** 3155595
- **Project number:** 5R01AR027940-11
- **Recipient organization:** UNIVERSITY OF TENNESSEE HEALTH SCI CTR
- **SAM.gov UEI:** X1M1PN3KG3E7
- **Principal Investigator:** DONALD J DONALDSON
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 1993
- **Award amount:** $154,832
- **Award type:** 5
- **Project period:** 1982-04-01 → 1994-11-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/3155595

## Citation

> US National Institutes of Health, RePORTER application 3155595, CELL-SUBSTRATE INTERACTIONS IN EPIDERMAL CELL MIGRATION (5R01AR027940-11). Retrieved via MORVS 2026-10-24 from https://api.morvs.ai/grant/nih/3155595. Licensed CC0.

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