System wide characterisation of cell adhesion in reproduction.

Cell recognition and adhesion are critical to all developmental processes. A paradigm of such is the apposition and attachment of the blastocyst to surface epithelium of endometrium to initiate developmental processes in the establishment of pregnancy. Ligands and their receptors important to this process have been described, none more so that the mucin muc-1 which in mouse gene knock-out studies has been demonstrated to be a vital component in influencing embryo adhesion to surface epithelium of the endometrium. Extension of such work to human implantation relies upon (a) characterisation of muc-1 expression in human endometrium and correlation with the opening and closing of the 'window of implantation'; (b) development of in vitro models in which to explore the role of muc-1 as a determinant of the adhesive properties of human endometrial epithelium. We have extensive experience of in vitro models of human endometrium established from clinical biopsies and their use to explore the molecular and cellular endocrinology of endometrial growth and differentiation. This project will exploit this expertise to investigate the effects of controlled expression of muc-1 upon surface properties of endometrial epithelium characterised by atomic force microscopy which provide both topographical and adhesion force data. Expression of muc-1 will be induced in response to progesterone or transfection of muc-1 cDNA; repression of muc-1 expression will be achieved by siRNA. Such cells will be analysed by AFM using cantilever tips functionalised either with isolated trophoblast cells or ligands that recognise components of muc-1 known to be presented at the cell surface of epithelium. Integration of this data will provide novel information on the contribution of muc-1 to adhesion properties at the surface of human endometrium. The project is in partnership with Veeco Instruments Ltd and formalises the relationship developed with them recenty. We also have agreement with the Science and Technology Facilities Council to exploit their novel technology in developing cantilever arrays; this relationship will be developed in the course of this project.