The human brain consists of intricately connected neurons forming functional circuits encoding memory, emotions and cognition. A neuron’s polarized organisation into axons and dendrites underlies its ability to receive, process and transmit information. To establish and maintain such a highly complex organisation, cytoskeletal motor proteins transport basic building blocks to either axons or dendrites. Several members of the kinesin motor family are selectively moving into axons, presumably guided by the specific organisation of the neuronal microtubule cytoskeleton. However, how different aspects of microtubule network organisation, such as filament length, orientation and modifications, contribute to selective transport of these kinesins is poorly understood. Therefore the aim of this project is to determine the connection between neuronal microtubule organisation and polarized transport of different kinesins. We will map the three-dimensional microtubule network at nanoscale resolution (objective 1) and decipher the intracellular transport driven by kinesins with optogenetics (objective 2). The integrated and quantitative knowledge provided by this project will pave the way towards understanding polarized transport in health and neurodegenerative diseases.