Subprojeto 16 – Mechanisms of cell signaling during adhesion, differentiation and cell death: identification of signaling pathways and association with physiological and metabolic aspects of living cells

Participantes: Carmen V. Ferreira; William Zambuzzi, Hernandes F Carvalho (IB-Unicamp)

Cell signaling mechanisms are usually fast events occuring in response to extracellular signals. We have been studying many biological models and pathways involving cell proliferation, differentiation and death by classical biochemical approaches. The purpose of this project is to integrate classical techniques such as Western blotting, by using antibodies against specific sites of phosphorylation (inhibitory or catalytic sites) for a series of pathways, with advanced microscope for the follow up of modifications of cell behavior, based on imaging components related to cell metabolism, such as NAD+/NADH ratio, lipids and membrane oscillations, in conjunct with calcium fluctuations in real time situations, by using FLIM, CARS and vital microscope measurements. Also, we will be concentrated on a model of pre-osteoblast cell adhesion and differentiation in response to 2D and 3D cultures.