EFucci an atlas of proliferation to reduce mouse usage in embryonic proliferation assays

Lead Research Organisation: University of Edinburgh
Department Name: MRC Human Genetics Unit

Abstract

Proliferation describes the process of growth and division of individual cells within a tissue in order to collectively increase its size during development or maintain its size once it is established. It is key to embryonic development and physically drives the growth of the embryo and the formation of tissue subtypes and organs. Regionalised changes in proliferation rates can help organs to develop and drive the determination of their shape or structure. For these reasons the study of proliferation is fundamental to almost all research groups who focus on embryonic development in the context of health and disease and also in many other contexts. There were over 3000 academic papers published in the field in 2013 alone. To mark dividing cells mice can be administered substances that integrate into their DNA during the cell division process, these animals can then be killed and the pattern of cells positive for these markers can be investigated. This experimental approach involves multiple injections and/or the surgical implantation of devices to slowly release these substances. We have developed an alternative mouse model to label dividing cells that does not require any surgical interventions. The purpose of this grant is to use this new model to create an atlas of embryonic development that allows research groups to better design and plan their proliferation assays and so reduce the number of mice they would require to carry out their studies. The atlas will also help to promote the use of our alternative model thus reducing the number of surgical procedures required to carry out the experiments.

Technical Summary

The study of proliferation is key to the characterisation of many models of human disease especially in the developing embryo. There were over 3000 academic papers published in the field in 2013 alone. Often these studies use the incorporation of thymidine analogues into the DNA of dividing cells to identify sub-populations of cycling cells. This approach usually requires either multiple intraperitoneal injections of the thymidine analogue or the surgical implantation of mini osmotic pumps to slowly release the analogue. The Fucci system is an alternative to this approach that uses two genetically encoded biosensors of cell cycle phase to label cells and therefore does not require surgical intervention. We have developed an alternative mouse model that incorporates the Fucci probes into a single construct driven by a strong ubiquitously active promoter (R26Fucci2a). This has allowed us to reduce animal usage in our own experiments by abrogating the need to use two mouse lines, one expressing each Fucci probe. #The atlas will also help to promote the use of our alternative R26Fucci2 model thus reducing the number of surgical procedures required to carry out the experiments.

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