Research Grants

Award Year 2004

TORRES Miguel
SHARPE James
Computerized 4-Dimensional Clonal Analysis of the Developing Mouse Limb

Co-ordination of the various complex processes that make up embryo development:gene regulation, cell-cell interactions, genetic networks, pattern formation, tissue mechanics and many others, is one of the remaining frontiers in biological science. Current tools for understanding these systems such as molecular biology, micro-manipulation and in-situ staining techniques, give us access to a wealth of information. However, techniques for following organogenesis at the level of individual cells are still limited. The aim of this project is to combine and further develop a number of recent technologies to trace the fates of individual cells within the developing vertebrate limb bud: (1) Molecular and genetic biology tools will be used to generate region-, tissue- and time-specific wild type and mutant labelled cell clones, so that comprehensive 3D maps of organogenesis can be compiled at a cellular level through Optical Projection Tomography (OPT)1 analysis (2) A new laser-scanning version of the OPT approach will be developed and applied to the analysis of labelled cell clones in living cultured mouse limbs. This will be the first time that such a high-resolution technique has been used to monitor the movements of fluorescently-labelled cells in live tissue culture. (3) A computational 4D framework will be programmed incorporating theoretical concepts for interpretation of clone distributions and prediction of growth dynamics. Together, these three tools will allow us to create a detailed 4D description of limb development at the cellular level. The end point of the whole project thus will be an understanding never before achieved for a developing vertebrate organ, namely a computerised 4D description which captures the dynamics of all tissue movements. This integrated approach will ultimately be applicable to understanding the development of any vertebrate organ and other processes such as adult organ regeneration or tumour formation. By analogy to the sequencing of the human genome, we can see that this will be both an end-point and a beginning: while it will immediately provide us with a far deeper understanding of cell fate, tissue dynamics and the basic processes that combine to allow limb development, it will also act as a new framework for the future within which scientists will be able to explore the genetic control of these complex processes.