Morphogenesis of the elastic fibre : a study in bovine ligamentum nuchae and human foetal aorta

During embryogenesis, the inherited information on the spatial arrangement of a particular tissue results in the deposition of extracellular matrix components. The overall tissue architecture is the responsibility of the embryonic cells which use the physico-chemical properties of the synthesised matrix components to assure local stability and form. Trelstad and Birk (1984) have postulated that embryonic cells are inherently polarised in both structure and function and that the quality, quantity and orientation of the extracellular matrix is under cellular control. They have shown that the cellular polarity and matrix patterning in embryonic chick tendon and corneal matrix is very similar to that proposed in this thesis for the morphogenesis of the elastic fibre. During matrix secretion, epithelial cells in the cornea are observed to undergo a reversal of their polarity with up to 80% of the cells relocating their Golgi to the basal cell pole. Discrete packets of collagen are then transported to the cell surface where the vacuoles are brought into register with the orientation of the underlying matrix by the alignment of ordered filaments and/or microtubules in the basal cell cytoplasm. The fact that elastin appears in the extracellular matrix at a later stage in development than most other connective tissue macromolecules suggests that elastogenic cells require a highly structured or 'mature' extracellular matrix upon which to organise the complex elastic fibre and that only late in gestation is the matrix suitable for the fibre organisation. Accordingly, a component of the extracellular matrix such as the microfibrils may provide the information that signals the appropriate time for elastin synthesis and secretion. This view is supported by the cellular polarity and the presence of both secretory vesicles and aligned microtubules (Chapter Two) observed when elastin-producing cells are in close proximity to the microfibrillar component of developing elastic fibres. The belief that elastin-associated microfibrils play a crucial role in the morphogenesis of the elastic fibre has until recently been based primarily on electron microscopic studies of the development of elastic tissue. In Chapter Three, purified 35k-GP was shown by immunoblotting and immunochemical techniques to be a constituent of elastin-associated microfibrils in developing elastic tissues within the ear, skin, aorta and ligamentum nuchae of the foetal calf. No binding of anti-35k-GP was detected in adult bovine ligamentum nuchae, aorta or ear which suggests that this glycoprotein plays an important but transient role in the development of elastic tissue. It is proposed that the role of 35k-GP in the function of microfibrils is to induce the adhesion and morphogenetic movements of elastogenic cells during elastic fibre formation. The preliminary results described in Chapter Four support this view and we now plan to undertake a more detailed study with the following objectives in mind:- 1.(a)To examine by immunofluoresence the involvement of cytoskeletal structures such as microtubules and microfilaraents in the interaction of elastogenic cells with 35k-GP. (b)To determine the type and duration of the 35k-GP induced adhesion by time-lapse video interference reflection microscopy to discover whether a substrate specific focal contact is occuring resulting in cytoskeletal reorganisation and ultimately in the synthesis and extracellular deposition of elastin. 2. A study of the ability of 35k-GP fragments to induce cell adhesion and spreading. This will allow the elucidation of a possible specific cell-binding region of the molecule. 3. An investigation into the identity of a specific cell receptor, the presence of which appears essential to the primary recognition event in the binding of 35k-GP and the possible existence of additional cell surface molecules to the cell-substratum attachment mechanism. 4. In conjuction with the above, a gene library from human foetal aortic smooth muscle cells is being created with the intention of developing cDNA probes to 35k-GP specific mRNA from this tissue. (Abstract shortened by ProQuest.