In vitro and in vivo tracing of fluorescently tagged cell types in the lung

Background:

Lung development process encompass various cellular processes including cellular interactions like alveolarisation, septa formation, differentiation and trans differentiation processes. Similar to this, some lung diseases like pulmonary fibrosis are characterized by tissue remodeling processes.

The aim of the first project is to visualize these interaction processes by specific fluorescent labelling of three different cell types by single plasmid containing fluorochromes under control of the respective cell type specific promotors. The triple-coloured plasmid of this project is finished and is now in the in vitro control.

The aim of the second project is the visualisation of transdifferentiation processes, using the inducible Cre/loxP, – FLP/FRT systems and overlapping fluorescences to specifically mark different celltypes which undergo these processes. For this project one construct, carrying the reporter genes and loxP- and FRT-sites is finished and successfully tested in vitro.

Outlook:

The further work of the first project will focus on the completion of the in vitro studies, validating the specific expression in different cell types as well as some functional studies to visualize epithelial mesenchymal transitions in vitro using TGFß. The aim is the creation of transgenic mice for in vivo analysis of tissue specific labeling which will facilitate cell specific sorting, in vivo imaging and stereological quantification of structural parameters.

The aim of the second project is the generation of a transgenic mouse line. Additionally the work will focus on the creation of the second mouse line, thereby concentrating on the design of an inducible, generalized FLP mouse line. The aim is to introduce this system as a general tool for visualization of transdifferentiation processes. It will be used for the in vivo investigation of transdifferentiation processes in lung development and lung diseases.