Secreted Proteins from Endothelial cells for Endocrine Development (SPEED)

The precise sequence of events during regeneration is difficult to asses using traditional stereological 2D analyses because they rely on certain assumptions, which might be unfounded. We propose to use an optical method for 3D projection tomography based reconstruction of the entire organ, which allows an accurate representation of islet distribution and volume. This method will be used to characterize b-cell regeneration following PDL. Furthermore, we will include 3D imaging of recruited inflammatory cells and reorganization of blood vessel, either by antibody mediated staining protocols or by lectin injections in the heart.
Finally, we propose to use the PDL model to characterize the entire proteome in three distinct sorted cell populations representing ductal progenitors, endocrine precursos and b-cells. We will use genetic models where cells can be FACS sorted and hereby purified based on GFP expression. The objective is to identify activated pathways using bioinformatics on the obtained protein expression data.