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C Ockleford

Publications and source records attributed to C Ockleford.

6 recordsLinked to original sources

Use of green fluorescent protein in visualisation of pneumococcal invasion of broncho-epithelial cells in vivo.

The pneumococcus is the principle cause of bacterial pneumonia and also a major cause of bacterial meningitis. The mechanisms and sites of pneumococcal adherence and invasion of the respiratory tract in vivo are not clear however. We have made pneumococci expressing green fluorescent protein (GFP) and used it to trace pneumococcal adherence and invasion in vivo. By using GFP pneumococci we have shown bacterial adherence and invasion of broncho-epithelial cells in vivo by 4 h post-infection, with increases in pneumococcal invasiveness by 24 h. Using confocal image analysis we have shown varying levels of pneumococcal penetration and internalisation into host cells, as well as translocation through epithelial layers. To our knowledge this is the first report of pneumococcal invasion and cellular translocation in vivo.

Animals↗

Regions responsible for organizing the microtubular cytoskeletons of extra-embryonic membranes.

We have identified putative microtubule-organising centres in whole mounts and frozen sections of full term human placenta, amniochorion and first trimester chorionic villus tissue by indirect immunofluorescence confocal laser scanning microscopy. These sites may perform a crucial executive role in the morphogenesis of human extra-embryonic membranes, and also may establish polarity in the amniotic epithelium and mediate the branching pattern of growth essential to the production of the chorionic villus tree.

Amnion↗

Micro-trabeculae, macro-plaques or mini-basement membranes in human term fetal membranes?

Immunocytochemical confocal laser scanning microscopy and ultrastructural analysis, including immunoelectron microscopy, reveals the distribution of structures in human term amniochorion similar in some respects to basement membranes but with unusually restricted dimensions. On the basis of their immunoreactivity, these trabecular structures, found on the fibroblast layer side of the spongy layer of human term amniochorion and adjacent reticular layer, have been shown to contain type IV collagen, laminin, and nidogen. The origin of these components may be from primitive epithelial structures which pumped fluid into the lakes that eventually coalesced to form the extraembryonic coelom separating the extraembryonic somatic mesoderm from the extraembryonic splanchnic mesoderm. Such a theory of their origin might link them with the mysterious 'cellular layer', a single-cell-thick layer of cells which is usually no longer present in fetal membranes at term. The similarity in composition but not in size of these structures to anchoring plaques for type VII collagen is possible support for the view that these structures are integrators of extracellular matrix polymeric proteins. The 'pseudobasement' membrane associated with the trophoblast layer, on investigation, appears to be typical by six criteria. 'Coiled' fibrous structures in the extracellular matrix of the spongy layer may aid adjustments under tension at this shear surface by a detachable 'Velcro' or 'two spring' fastening system. The coils are rich in fibronectin. The suggestion is made that the compact layer is a giant lamina reticularis associated with the amniotic epithelial basement membrane.

Basement Membrane↗

Confocal and conventional immunofluorescence and ultrastructural localisation of intracellular strength-giving components of human amniochorion.

Key cytoskeletal polypeptides of human fetal membranes have been localised at subcellular level using confocal and conventional indirect immunofluorescence microscopy. Correlation with electron microscope data has allowed us to examine how cellular compartments of this multilaminar tissue maintain their mechanical integrity until the time of membrane rupture at parturition. Evidence is presented for myofibroblastic characteristics of cells in both the fibroblast and reticular layers which may therefore have tension-generating, position-adjustment and wound-healing roles in the amniochorion. Desmin and vimentin are coexpressed in these cells, but a small localised population of cells in the fibroblast layer contains vimentin alone. An interaction of cytokeratin filaments with nuclei and desmosomes of amniotic epithelium in vivo is demonstrated, indicating that nuclei of cells of ectodermal origin are integrated into a mechanical structure extending throughout the tissue as a whole. Cells of the basal 1 or 2 layers of trophoblast have been shown to have a more extensive and better integrated cytoskeletal organisation than those overlying and forming the boundary with decidua. Structures within the trophoblast, identified previously as degenerate villi, contain cells with intermediate filaments with similar immunofluorescence properties to those of the neighbouring reticular layer and thus may represent papillae that prevent shearing at this interface.

Amnion↗

Hydatidiform mole: an ultrastructural analysis of syncytiotrophoblast surface organization.

The scanning ultrastructural examination of a series of 31 hydatidiform mole and 12 healthy placental specimens of similar gestational age has revealed a variety of surface architectures more common in molar tissue. Characteristic paddle-shaped sprouts, ridging of the syncytial maternal oriented surface and microgibbosities are described. These structures are explicable in terms of organellar hyperplasia of cortical cytoskeletal elements found in healthy tissue. Specific morphological evidence of involvement of these elements in a condition where aberrant growth control leads to the characteristic trophoblastic hyperplasia is a further indication that cytoskeletal elements may mediate transformation. An increase in resolution obtained over previous scanning electron microscope studies has allowed the description of detailed features such as 'caveolar collars' on the maternal oriented healthy and molar trophoblast surfaces. These observations are of relevance to understanding the mechanisms of several cell physiological processes, including transepithelial transport. New observations of a reticular organization in the surface layer of molar trophoblast indicate that a syncytioskeletal layer, with organization resembling that previously described in healthy chorionic villi, is also present in molar villi.

Chorionic Villi↗