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Title: Molecular basis of fibrillin-rich microfibrils and cellular interactions
Author: Bernard, Sarah Elizabeth
ISNI:       0000 0001 3461 5749
Awarding Body: University of Manchester : University of Manchester
Current Institution: University of Manchester
Date of Award: 2002
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Fibrillin-rich microfibrils are key structural elements of both non-elastic and elastic dynamic connective tissues. Fibrillin-rich microfibrils in situ are often found in close proximity to cells. Fibrillins contain ROD putative cell recognition sequences and fibrillin-l and fibrillin-2 peptides containing these sequences have been shown to interact with cells via avf33 and f31 integrin receptors. This study is concerned with elucidating the molecular basis of cell-matrix interactions of purified assembled fibrillin-rich microfibrils and skin cells. Fibrillin-rich micro fibrils have been isolated and purified from neonatal bovine aortae, and deemed to be suitable as ligands for cell based assays. In addition, recombinant human fibrillin-l constructs produced in mammalian cells were employed as ligands in cell attachment and cell spreading assays to localise the site of cellular interaction from a large intact polymer to a few domains in the fibrillin-l molecule. Human dermal fibroblasts and HaCaT cells have been shown to attach and spread on intact fibrillin-rich microfibrils and recombinant fibrillin-l ligands. This cell attachment has been shown to be dependent on the ROD sequence in the fourth TB module of fibrillin. Furthermore, avf33 and CXSf31 integrins have been identified in mediating human dermal fibroblast attachment, whilst CXSf31integrin has been implicated in mediating HaCaT cell attachment to fibrillin. Immunofluorescent microscopy studies have determined the time scale of integrin receptor and paxillin expression in human dermal fibroblasts plated on fibrillin-rich microfibrils. Human dermal fibroblast attachment is initially mediated by CXs~1 but, av~3 is also involved in the attachment Furthermore, paxillin expression indicates the formation of focal adhesions over the time scale of human dermal fibroblasts attaching to fibrillin-rich microfibrils. Cell signalling assays did not detect tyrosine phosphorylation of focal adhesion proteins over the time scale of human dermal fibroblast attachment and spreading on fibrillin-rich microfibrils. These results are an important addition to the advancement of deciphering the critical role fibrillin-rich microfibrils play in the regulation and organisation of specific tissue extracellular matrices. This work was supported by the Biotechnology and Biological Sciences Research Council (BBSRC), UK.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available