Maternal communication with gametes and embryos influences a broad range of events crucial to pregnancy. Events such as final maturation of gametes, gamete transport, fertilization, early embryonic development and development of foetus to term, are all dependant upon the relay of appropriate molecular signals between the mother, gametes and embryos. This signalling is temporally and spatially regulated, involving complex interactions. Disturbances in maternal communication with gametes and embryos can influence the outcome of pregnancy. Effects range from those that are immediately obvious, such as spontaneous miscarriage (due to inappropriate hormonal signalling), to more subtle consequences that may not become apparent until offspring reach adulthood ('foetal origins' hypothesis). Current knowledge of the factors and mechanisms involved in maternal communication with gametes and embryos is limited to only a few individual pathways. There is a need for a holistic view of all actions and interactions taking place during this crosstalk between the gametes, embryos and the female reproductive tract. Applying high-throughput genomic and proteomic analysis tools and systems biology approaches, together with mathematical modelling would allow construction of an in silico model for the temporal sequence of events involved. Ultimately this will help identify different dimensions of maternal communication with gametes and embryos in health and disease.

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The use of stem cells for generating cell types suitable for therapy is dependent on understanding the mechanisms, and identifying biomarkers, that control cell fate into different lineages. In this study, we aimed to characterize the nuclear protein dynamics of NTERA-2 cells undergoing retinoic acid-induced differentiation. We focused specifically on the first six days of differentiation, to provide insight into the earliest differentiation events, and employed techniques to specifically monitor the nuclear proteome. Well-characterized gene expression markers were used to precisely stage cell differentiation across the experimental time course. A combination of the novel iTRAQ and ExacTag labeling technologies, together with LC-ESI tandem mass spectrometry, were then used to accurately measure nuclear protein expression changes occurring within these differentiation-staged cells. We report proteins that showed significantly altered expression over the first 6 days of differentiation. Extensive bioinformatic analysis was undertaken, resulting in the construction of a novel interactome network, which revealed the temporal dynamics of the nuclear protein network in the context of neuronal differentiation.

The antigenic profile of human embryonic stem (ES) and embryonal carcinoma (EC) cells has served as a key element of their characterization, with a common panel of surface and intracellular markers now widely used. Such markers have been used to identify cells within the 'undifferentiated state', yet it appears that this categorization may be an oversimplification, because a number of sub-states appear to exist within this state. To increase the resolution of the undifferentiated state, we have generated eight novel monoclonal antibodies, all capable of recognizing undifferentiated human ES and EC cells, and herein describe their characterization. The reactivity of these antibodies against a range of cell lines is reported, as well as their developmental regulation, basic biochemistry and reactivity in immunohistochemistry of testicular germ cell tumours. Our data reveal a range of reactivity for all antibodies against both ES and EC cells, suggesting that these markers will afford recognition of unique sub-states within the undifferentiated stem cell compartment.

The functional reprogramming of a differentiated cell to a pluripotent state presents potential beneficial applications in regenerative medicine. We report here the proteomic profile of 293T epithelial cells reprogrammed to a pluripotent state using undifferentiated embryonal carcinoma (NCCIT) cellular extracts. 293T cells were reversibly permeabilized with streptolysin O, incubated in an extract of NCCIT cells or a control extract of 293T cells for 1 h, resealed with CaCl 2 , and cultured. OCT4 and SOX2 gene expression were up-regulated in NCCIT extract-treated cells relative to control cells, whereas there was no alteration in DNMT3B gene expression. Thirty percent of NC-CIT extract-treated cells were positive for SSEA-4, and karyotyping confirmed their 293T origin, excluding the possibility of contamination from NCCIT cells. Two-dimensional PAGE revealed ϳ400 protein spots for each cell type studied. At least 10 protein spots in the proteome of NCCIT extract-treated cells had an expression profile similar to that of NCCIT and remained unaltered in control cells. Using tandem mass spectrometry, we identified these proteins, which include 78-kDa glucose-regulated protein precursor and tropomyosin ␣-3 chain. This investigation provides the first evidence that proteins are altered in a specific manner in NCCIT extract-treated cells.