THE CA2+ SIGNAL REGULATES MANY DIFFERENT CELLULAR FUNCTIONS, SO CORRECT INTRACELLULAR CALCIUM HOMEOSTASIS REQUIRES ITS SUBCELLULAR COMPARTMENTALISATION AND THE GENERATION OF MICRODOMAINS WITH DIFFERENT CA2+ CONCENTRATIONS. THE TRANSPORT OF CA2+ THROUGH INTRACELLULAR ORGANULS (ENDOPLASMICO-RE-, MITOCHONDRIA, NUCLEO, GOLGI -GO) IS ESSENTIAL IN THE GENESIS OF MICRODOMAINS. IN ADDITION, THE CONCENTRATION OF CA2+ WITHIN ORGANULOS IS ESSENTIAL FOR MANY PHYSIOLOGICAL FUNCTIONS (PROTEIN PROCESSING, BREATHING, GENIC EXPRESSION, APOPTOSIS). IN FACT, CA2+ DISHOMEOSTASIS APPEARS TO PLAY A VERY IMPORTANT ROLE IN THE PATHOGENESIS OF NEURODEGENERATIVE DISEASES. IN THE PREVIOUS PROJECT WE HAVE DEVELOPED A NEW FAMILY OF CA2+ FLUORESCENT INDICATORS, WHICH WE CALL GAP (FROM GFP-AEQUORIN-PROTEIN), AND THAT CAN ADDRESS THE MATRIX OF INTRACELLULAR ORGANULOS OF LIVE CELLS (NORMAL OR PATHOLOGICAL!) AND MONITOR IN REAL TIME THE CONCENTRATION OF CA2+ INSIDE AT REST AND/OR DURING PHYSIOLOGICAL ACTIVITIES. IS LCREO, IN ADDITION, A TRANSGENIC RATON OF NORMAL PHENOTYPE AND EXPRESSING FUNCTIONAL GAP IN ITS RE (PNAS 111:2584-89; 2014), AND WHICH HAS HAD A REMARKABLE IMPACT AND ACCEPTANCE. IN THIS PROJECT WE INTEND TO EXPLOIT THIS TOOL BY FOCUSING ON RE AND GO, WHOSE CA2+ CONTENT IS ESSENTIAL FOR THE PROCESSING OF PROTEINS OF VIA SECRETORA. They PROPONATE EXPERIMENTS that were grouped into three OBJECTIVES, GENERALS INCLUDING STRUCTURAL, PHYSIOLOGICAL AND physiopathological STUDIES:_x000D_ 1. NEW METHODOLOGICAL DEVELOPMENTS AND IMPROVEMENTS OF THE GAP. IT IS PROPOSED TO REDUCE THEIR AFFINITY FOR MUTAGENESIS OF RU REGIONS TO IMPROVE SENSITIVITY IN RE AND GO, TO MANUFACTURE VARIANTS FOR SELECTIVE EXPRESSION IN SPECIFIC CELL TYPES (NEURONS, GLIAS, ETC.). IN COLLABORATION WITH OTHERS WE INTEND TO CARRY OUT STRUCTURAL AND MECANISTIC STUDIES AROUND THE GAP-CA2+ INTERACTION. Also IN COLABORATION, we intend to GENERE NEW TRANSGENICAL ANIMALS (NEW RATONES, MOSCAS AND GUSANS)._x000D_ 2) STUDY OF CA2+ HOMEOSTASIS IN RE AND GO AND RELATION WITH DIFFERENT physiologic FUNCTIONS. IN COLLABORATION WITH OTHER LABORATORIES WE PROPOSE A SEARCH FOR THE LEAKAGE CHANNEL (LEAK) BASED ON COMPARATIVE PHYSIOLOGY AND EVOLUTIONARY GENETICS, AND A STUDY OF THE ROLE OF ELECTRIC BALANCING OF CA2+ RELEASED FROM RE/GO IN NEURONS AND GLIAS. Collabrations are PLANT WITH OTHER GROUPS TO STUDY THE PARTICIPATION OF THESE ORGANISATIONS IN varietal physiologic functions (secretion of HORMONAS, GLIAL oscillations, SENSORIAL CELL ACTIVATION, ETC)._x000D_ 3) PROPONE, finally, to be able to establish the potentials of CA2+ HOMEOSTASIS IN RE and GO IN THE Aging AND IN NEURODEGENERATIVE DISEASES MODELS. WE WANT TO COMPARE CA2+ LEVELS IN D AND GO NORMAL AND SICK CELLS (INCLUDING NEURONS AND GLIAS!). All PROJECTS IN COLLABORATION that will include models of ageing, ALZHEIMER (Preseniline TRANSGENIC TRIPLE, B-AMILOIDE and TAU PRECURSER), PARKINSON AND HUNTINGTON, INCLUDING TWO ULTIMES MODEL IN MODEL CELLS WITH INDUCED pluripotence (IPS) and DIFFERENTIALS TO DOPAminergical NEURONAS OR STRIATAL PROJECTION NUERONAS._x000D_ _x000D_ There is no way to inflate, NI IN THE INVESTIGATION TEAM NI IN THE WORKING TEAM TO MY Fellows PREDOCTORAL, PALOMA NAVAS NAVARRO AND MACARENA RODRIGUEZ PRADOS. I NOTE THAT THEY ARE INVOLVED IN THE PROJECT.