The precise connection between neurons during development is one of the most accurate and regulated processes which contributes to the physiological function of neuronal circuits within the brain. Eph/ephrins play a crucial role in the development of neural connectivity. During early development, and across adulthood, Eph/ephrins support spatiotemporal, topological as well as specific clues, and deregulation of its function is observed in many neurodegenerative diseases. Up to now, the precise domains/motifs involved in Eph functions are not fully dilucidated. Here, we assessed cultured cells expressing fluorescent EphA3-WT and truncated vectors in order to determine vesicular parameters of axonal transport by live imaging recording movies like kymographs, segmental velocities, etc. Preliminary results showed that WT had mostly a vesicular mobile phenotype. Meanwhile, ligand binding truncated versions showed both an increased distribution in the plasma membrane region coupled with increased vesicle density and mobility compared to the WT. Furthermore, we will examine Eph transport in human glutamatergic neurons derived from isogenic-control and mutated tau iPSC as a model of neurodegenerative diseases. We propose that understanding the requirements for Eph/ephrins transport and distribution is key to unravel brain wiring mechanisms providing knowledge to comprehend the abnormalities that are associated with synapse collapse during neurodegenerative diseases.