Quantitative clonal analysis and modelling of retinogenesis in zebrafish (360G-Wellcome-105355_Z_14_Z)

In order to understand how organs achieve their specific sizes and cell compositions, it is critical to have quantitative details on the proliferation and fate choices of the progenitor cells through developmental time. The zebrafish retina is an excellent system to study this process in the central nervous system of a vertebrate, and to date we have quantitative models that do a reasonable job at predicting how the zebrafish retina is generated. There are still some uncertainties and non-fixed parameters in these models and the models are not yet fully accurate. The project here is to obtain a great deal more data so that we can generate more accurate and constrained models. Key Goal 1: To optimize a protocol for imaging the whole zebrafish retina so that it will be possible to follow the full clonogenesis and differentiation of retinal progenitor cells (RPCs), from 24 hours post fertilization (hpf) to 72 hpf when central retinal development is complete. Key Goal 2: To generate a computational image analysis based toolset to track the lineages resulting from RPCs, as well as their movement in three dimensions. Key Goal 3: To extend the current model the RPC fate decision-making process so that it more fully and accurately explains clone size and cell composition. Key Goal 4: To develop a method for in vitro clonogenesis and thereby to study the development of zebrafish RPCs plated at clonal densities