Periodic Report Summary - PERRY/MII/TG (Transgenic models of mammalian meiotic exit) The objectives were to generate mouse transgenic lines with which to dissect the egg-to-embryo transition (metaphase II, mII, exit) immediately after fertilisation. The transgenic lines are intended to express markers of key processes during this transition, including fluorescent proteins fused to cytoskeletal components, cell cycle regulators and chromatin components; these reagents either do not yet exist, are sub-optimally configured or are not widely available. To this end, and in the transposition of my laboratory from Japan to England, IRG support has been indispensible.The project is currently at least on schedule, if not slightly ahead of it. This is notwithstanding what in hindsight was a somewhat ambitious programme; relocating a mouse embryology group from Japan to England alone is a major task. Owing to a delay in confirming ethical approval, the project did not commence until September 2010, 5 months after the contractual starting date (April, 2010), which is when I arrived in England. Since commencement we have successfully established a micromanipulation laboratory in the University of Bath Animal Facility. Equipment has been installed following repair where necessary. A breeding programme has been initiated to provide the standard mouse stock strains for our work. All necessary ethical and other permissions, including licenses mandated by the Animals (Scientific Procedures) Act, 1986 for the proposed work have been obtained (PILs for all workers; PPL No. 30/2795, held by Dr A.C. Perry). Embryo culture and transfer have been established and optimised at Bath. In achieving this goal, we have established perhaps the first and only piezo-actuated microinjection laboratory in England. This was demonstrated by proof-of-principle experiments in June/July 2011, in which we produced what are probably England's first nuclear transfer cloned mice; the surviving clone, Yuami, can be viewed in 'Interim report: Progress and achievements' available at http://www.egg2embryo.com/.We have initiated a programme of transgenesis and are pleased to report the production of founder lines corresponding to four constructs, including the de novo generation of lines ubiquitously-expressing eGFP. We are now characterising and expanding these lines for use as reporters in experiments to dissect processes in very early development, in particular, key mechanisms in genomic reprogramming after gamete union in fertilisation. Our original plan envisioned first generating transgenic lines for the study of cell cycle and cytokinetic regulation but we have instead first generated lines expressing chromatin markers, as these have greater and broader utility. The over-arching goal is a mechanistic understanding of the regulation of cellular potency; we consider fertilisation to be the most extreme programmed cellular potency switch, so understanding it well will be necessary for any model hoping to explain the genesis of induced pluripotent stem (iPS) cells and cancer. Clearly, in as much as they contribute to such an understanding, the present work has major socio-economic implications for biomedical progress, and the IRG proposal has enabled us to work to this end at home and by consolidating and extending collabourations within the EU and further afield. Other report summaries Periodic Report Summary 2 - PERRY/MII/TG (Transgenic models of mammalian meiotic exit)
