The biological potential of extracellular vesicles (EVs) arises from their ability to transport and deliver cargo within and across cells. While widely accepted as a cell-cell communication vehicle, the dynamic range of these vesicles during brain development remains largely unexplored. Knowing this, we aimed to identify novel avenues of cellular communication and unravel the contribution of EV cargo to brain development. Leveraging a multi-omics approach, we profiled the miRNA, RNA, and proteomic contents of EVs spanning embryonic to juvenile states of mouse brain development. With this approach, we were able to define the developmental trajectories of these EV cargoes in the brain. Interestingly, we detected several proteins encoded by genes associated with neurodevelopmental disorders. We selected the ribonucleoprotein HNRNPU for further validation and functional studies due to its strong association with neurodevelopmental disorders as well as its molecular gene pleiotropy. Using CRISPR-Cas9, we developed a mouse model of tagged HNRNPU to confirm the presence of this protein in EVs using transmission electron microscopy, as well as to identify the interactome of HNRNPU in brain EVs. To identify the effects of Hnrnpu haploinsufficiency in brain development, we assessed the brain transcriptome of Hnrnpu+/- mice at E16.5 and identified differential expressed genes converging to processes related to RNA processing and brain development. Altogether, we are building evidence supporting the role of EV transport of an RNA-binding protein in post-transcriptional regulation of gene expression during brain development.