Lifeboat Foundation

Presynapses locally recycle synaptic vesicles to efficiently communicate information. During use and recycling, proteins on the surface of synaptic vesicles break down and become less efficient. In order to maintain efficient presynaptic function and accommodate protein breakdown, new proteins are regularly produced in the soma and trafficked to presynaptic locations where they replace older protein-carrying vesicles. Maintaining a balance of new proteins and older proteins is thus essential for presynaptic maintenance and plasticity. While protein production and turnover have been extensively studied, it is still unclear how older synaptic vesicles are trafficked back to the soma for recycling in order to maintain balance. In the present study, we use a combination of fluorescence microscopy, hippocampal cell cultures, and computational analyses to determine the mechanisms that mediate older synaptic vesicle trafficking back to the soma. We show that synaptic vesicles, which have recently undergone exocytosis, can differentially utilize either the microtubule or the actin cytoskeleton networks. We show that axonally trafficked vesicles traveling with higher speeds utilize the microtubule network and are less likely to be captured by presynapses, while slower vesicles utilize the actin network and are more likely to be captured by presynapses. We also show that retrograde-driven vesicles are less likely to be captured by a neighboring presynapse than anterograde-driven vesicles. We show that the loss of synaptic vesicle with bound molecular motor myosin V is the mechanism that differentiates whether vesicles will utilize the microtubule or actin networks. Finally, we present a theoretical framework of how our experimentally observed retrograde vesicle trafficking bias maintains the balance with previously observed rates of new vesicle trafficking from the soma.

Cytoskeleton-based trafficking mechanics have long been explored because of their essential role in neuronal function and maintenance (Westrum et al., 1983; Okada et al., 1995; Sorra et al., 2006; Perlson and Holzbaur, 2007; Tao-Cheng, 2007; Hirokawa et al., 2009; Staras and Branco, 2010; Tang et al., 2013; Wu et al., 2013; Maeder et al., 2014; Guedes-Dias et al., 2019; Gramlich et al., 2021; Watson et al., 2023). Protein trafficking via cytoskeleton transport is essential for synaptogenesis (Perlson and Holzbaur, 2007; Santos et al., 2009; Klassen et al., 2010; Wu et al., 2013; Guedes-Dias et al., 2019; Guedes-Dias and Holzbaur, 2019; Kurshan and Shen, 2019; Watson et al., 2023) and to replace older proteins with newer proteins for efficient function (Cohen et al., 2013; Dörrbaum et al., 2018, 2020; Heo et al., 2018; Truckenbrodt et al., 2018; Jähne et al., 2021; Watson et al., 2023).