CONICET | Buscador de Institutos y Recursos Humanos

CLONAL ANALYSIS OF STEM/PROGENITOR CELLS IN CHICKEN NEURAL RETINACindy Lorena Olmos Carreño (1°), Maria Figueres Oñate (2°), Mario Sanchez (2°), Gabriel Scicolone (1°), Laura López Mascaraque (2°)Filiaciones: 1° Instituto de Biología Celular y Neurociencias ?Profesor Eduardo De Robertis? (UBA-CONICET), Facultad de Medicina, UBA, Buenos Aires, Argentina., 2° Instituto Cajal-CSIC, Madrid, España, 3° , 4° .Clonal cell analysis defines the potential of single cells, allowing to decode neural heterogeneity of cell lineages and their clonal relationships. We adapted the mouse genetic tracing strategy UbC-StarTrack to a chick model. UbC-StarTrack is based on transfection of genes encoding fluorescent reporter proteins, six in the cytoplasm and six in the nucleus, driven by an ubiquitous promoter in PiggyBac-based vectors. This method produces inheritable marks that enable long-term in vivo cell tracing and attributes a unique color-code to single neural precursors, determining their differentiation potential and degree of dispersion. Once probed the accurate expression of these constructs in neurospheres obtained from dissociated cells of the chick retinal ciliary margin (CM) at 7 days of development (E7), UbC-StarTtrack mixture was co-electroporated into the retinal CM at E3.5. Labeled cell progenies were analyzed at different time points (2, 5 and 8-days postelectroporation). This allowed us to determine both, cell types originated from single cells and their clonal relationships within the retina. In conclusion: 1) UbC-StarTrack is valid in chicken model. 2) Cell clones formed columns extended between the inner and outer limitants of neural retina. 3) Cell clones displayed a large dispersion along the dorso-ventral axis but a limited dispersion by the anterior-posterior axis. 4) Different types of cells presented similar color combinations, revealing multipotency of some clonesCellular and Molecular Neurobiology