Final Report Summary - NEUROMOLANATOMY (The Molecular Anatomy of Neurons) The brain is nowadays the object of a number of extensive systematic studies that focus on seemingly all aspects of its morphology and function, from overall brain architecture to neuronal connections, neuronal morphology and gene expression. However, one basic aspect is as yet incompletely studied and understood: the molecular anatomy of the neuron, meaning the copy numbers and the spatial arrangement of the molecules within the neuronal cell. This cannot be addressed by gene expression or proteomics approaches, as they do not have sufficient spatial precision. Electron microscopy, ideal for unraveling the neuronal morphology, does not have sufficient protein labeling efficiency. This project fills this gap in our knowledge by a combination of super-resolution fluorescence microscopy, advanced fluorescence labelling techniques and advanced biochemistry tools such as quantitative mass spectrometry, thereby resulting in a complete 3D molecular view of the neuron. We determined the molecular organization of >100 major neuronal proteins, identifying their exact copy numbers and describing their position within the cell with nanoscale precision. We integrate the results into an in silico neuronal model that can be used for modelling functional neuronal parameters. Moreover, we have invented and optimized new assays for imaging the positions of the individual proteins with very high precision. At the same time, we have invented methods of imaging the neuronal metabolic organization with precision in both space and time. These tools also enable us to image neuronal lipids. This information, along with the copy numbers and the positions of the proteins, is now being integrated in a molecular picture of the neuron. Overall, our work generated the first ever quantitative molecular view of a cell – and of an extremely important cell, the neuron.
