Introduction
Although EVs are thought to be secreted by all cells, cell type-specific characterization of EV secretion is still rather limited. This is important, given the often large differences in gene expression profiles in cells of different origin and developmental stage. Indeed, parallel analysis of EV composition in different cell types and biofluids shows the discrete distribution of EV proteins, including commonly used markers, CD63, CD81 and CD9(Garcia-Martin, Brandao, Thomou, Altindis, & Kahn, 2022; Wiklander et al., 2018). This could be particularly relevant for highly differentiated cell types, such as neurons, that are known to display developmental, regional and functional specificity in gene expression, and that have adapted mechanisms of protein trafficking due to their highly complex morphologies. EVs were previously shown to mediate communication between neurons and non-neuronal cells(Bahrini, Song, Diez, & Hanayama, 2015; Men et al., 2019; Mukherjee et al., 2020), and have been implicated in the spreading of misfolded, aggregating proteins in neurodegenerative disease models(Asai et al., 2015; Lim et al., 2019; Sardar Sinha et al., 2018; Stuendl et al., 2016; Wang et al., 2017). Inter-neuronal transfer of EVs was also shown to be important for the regulation of synapse formation (Bahrini et al., 2015; Lee et al., 2018) and neurotransmission(Antoniou et al., 2023; Vilcaes, Chanaday, & Kavalali, 2021). Here, we evaluate EV secretion from primary neuron-enriched cultures and characterize the protein composition of neuronal-derived EVs using mass spectrometry.
Materials & Methods
List of antibodies
