Dynamic proteomics of mice hippocampus to study apoptosis

Apoptosis or physiological programmed cell death is a molecular process observed during neurodegenerative diseases (Epilepsies, Alzheimer…). The neurodegeneration leads to the reorganization of synapses and neurons in the CNS. Consequently, searches for early medication are a major challenge in Neuroscience. To this aim, VALAPODYN, a European Commission funded research network, develops functional genomics and proteomics related to the dynamics of molecular interaction networks (MIN). MIN modeling investigates protein-protein interactions and regulation networks. Using a model of induced hippocampal sclerosis associated with focal epilepsy in the mouse, dynamic expression analyses are conducted at different time points. Hippocampal samples were subjected on 2D-DIGE analysis which allows quantification of cyanine labelled proteins. Biological variation analysis yields about 1500 protein spots. These spots include 34 protein spots to be differentially regulated. The distribution is consistent with the literature showing that CNS proteins are mainly composed of acidic and neutral proteins. Several proteins have multiple protein spots likely resulting from different isoforms. Spots of interest will be digested with proteases and analysed using MALDI/TOF-TOF. Total proteins mixture will also be submitted to ESI-MS after HPLC separation as a means of improving both protein and proteome coverage by using complementary instruments.‘Validated Predictive Dynamic Model of Complex Intracellular Pathways Related to Cell Death and Survival