TEMPORAL PATTERNING OF NEURONAL PROGENITORS: FROM NEURAL DIVERSITY TO FUNCTIONAL...
DURING BRAIN DEVELOPMENT, PROGENITOR CELLS BALANCE PROLIFERATION AND DIFFERENTIATION TO GENERATE AN ENORMOUS DIVERSITY OF NEURONS IN THE CORRECT NUMBERS AND PROPORTIONS THAT EVENTUALLY INTEGRATE THE PROPER FUNCTIONAL CIRCUITS. ADD...
DURING BRAIN DEVELOPMENT, PROGENITOR CELLS BALANCE PROLIFERATION AND DIFFERENTIATION TO GENERATE AN ENORMOUS DIVERSITY OF NEURONS IN THE CORRECT NUMBERS AND PROPORTIONS THAT EVENTUALLY INTEGRATE THE PROPER FUNCTIONAL CIRCUITS. ADDRESSING HOW THIS OCCURS HAS BEEN TECHNICALLY CHALLENGING TO DATE. FOR INSTANCE, NO IN VITRO MODEL FULLY RECAPITULATES THIS IN VIVO PROCESS BECAUSE CELL CULTURES MISS THE 3D-TISSUE COMPLEXITY, AND BRAIN ORGANOIDS LACK THE TOPOLOGY OF THE BRAIN. ALTHOUGH SEVERAL REPORTS SHOWED THAT ORGANOIDS MAY CONTAIN THE DIFFERENT TYPES OF NEURONS, UP TO NOW THEY DO LACK THE PROPER NEURONAL SPATIAL ORGANIZATION.IN 4DNEURAMICS, WE WILL STUDY EMBRYONIC BRAINSTEM DEVELOPMENT IN ZEBRAFISH TO ADDRESS SOME OF THE MAIN UNSOLVED QUESTIONS IN DEVELOPMENTAL NEUROBIOLOGY AND REGENERATIVE RESEARCH: HOW DOES THE BRAIN GENERATE ITS ENORMOUS DIVERSITY OF NEURONS IN THE CORRECT NUMBERS AND PROPORTIONS? WHAT ARE THE MECHANISMS BEHIND? AND HOW ARE NEURAL CIRCUITS ASSEMBLED DURING EMBRYONIC DEVELOPMENT? ZEBRAFISH EMBRYONIC BRAINSTEM DEVELOPMENT IS A GOOD MODEL, AS IT REQUIRES BOTH DYNAMIC SELF-ORGANIZATION AND DRAMATIC MORPHOGENETIC CHANGES FOR ITS FINAL FUNCTIONAL OUTCOME. FURTHER, AND IMPORTANTLY, THE HINDBRAIN IS A HIGHLY CONSERVED BRAIN STRUCTURE IN VERTEBRATES.CRITICALLY, WE WILL INCORPORATE TIME AS A CRUCIAL BUT POORLY UNDERSTOOD FACTOR. THE GENETIC REQUIREMENTS FOR NEUROGENESIS HAVE BEEN ADDRESSED, AND THE IMPORTANCE OF BHLH FACTORS HAS BEEN REVEALED. NEVERTHELESS, INFORMATION ABOUT THE RELATIVE SPATIOTEMPORAL DISTRIBUTION OF NEURAL PROGENITOR POOLS UPON MORPHOGENESIS AND THEIR CLONAL RELATIONSHIPS REMAINS ELUSIVE DUE TO THE DEMANDS ASSOCIATED WITH THE IN VIVO IMAGING OF THE EMBRYONIC BRAIN. THIS STRUCTURE UNDERGOES EXTENSIVE MORPHOLOGICAL CHANGES WHILE CELLS PROLIFERATE, CHANGE SHAPE, AND DIFFERENTIATE. THEREFORE, RECONSTRUCTING UNAMBIGUOUSLY THE CELL LINEAGES IS CENTRAL TO THE UNDERSTANDING OF HOW THE PRECISE DIVERSITY OF CELL TYPES DEVELOPS TO GENERATE SPECIFIC CIRCUITS. HOWEVER, THIS CONSTITUTES A MAJOR CHALLENGE, AS IT REQUIRES SIMULTANEOUSLY TRACING THE LINEAGES WHILE RESOLVING THE KINETICS OF CELL PROLIFERATION. OUR OBJECTIVES AIM AT I) ELUCIDATING HOW CELL PROLIFERATION AND DIFFERENTIATION ARE INTERTWINED AND INFLUENCE TISSUE GROWTH AND CELL FATE, II) DISSECTING THE CONTRIBUTION OF DIFFERENT POOLS OF PROGENITORS TO SPECIFIC NEURONAL CIRCUITS, AND III) DECIPHERING THE GENE REGULATORY NETWORKS (GRN) OPERATING IN THE TEMPORAL PATTERNING OF SPECIFIC PROGENITOR CELL POPULATIONS TO CONTRIBUTE TO DISTINCT NEURONAL CIRCUITS. TO MIRROR RECONSTRUCTION OF THE DIVERSITY OF CELL LINEAGES, WE WILL MERGE INFORMATION FROM MORPHOGENESIS AND CELL FATE, GAINED BY THE USE OF COMPLEMENTARY STRATEGIES: THOSE USING POPULATION AVERAGE MEASURES THAT PROVIDE INFORMATION ON PROLIFERATION KINETICS AND CELL LINEAGE SPECIFICATION, AND THOSE COLLECTING INFORMATION AT THE CLONAL LEVEL PROVIDING INSIGHT INTO THE BEHAVIOR OF INDIVIDUAL CELLS IN CIRCUIT ASSEMBLY. THESE KINDS OF ANALYSES HAVE BEEN DIFFICULT TO ACHIEVE IN VERTEBRATE EMBRYOS DUE TO THE CHALLENGES OF DEEP-TISSUE IMAGING. HOWEVER, THE RECENT TECHNOLOGICAL INNOVATIONS IN FRONT-END MICROSCOPY ASSOCIATED WITH THE DEVELOPMENT OF ORIGINAL METHODOLOGIES AND TOOLS FOR THE IN VIVO MULTISCALE AND MULTIMODAL OBSERVATION, QUANTIFICATION, AND MULTILEVEL THEORETICAL MODELLING, WILL NOW ALLOW US TO DO A BIG LEAP FORWARD IN THE PREDICTIVE UNDERSTANDING OF BRAIN MORPHOGENESIS. INDBRAIN\MORPHOGENESIS\CELL LINEAGE\NEURAL CIRCUITS\NEURONAL DIFFERENTIATION\NEUROGENESIS\PROGENITORS\PRONEURAL GENES\CLONAL GROWTH\TISSUE SEGMENTATIONver más
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