Subdiffractional tracking of internalized molecules reveals heterogeneous motion states of synaptic vesicles

Merja Joensuu, Pranesh Padmanabhan, Nela Durisic, Adekunle T.D. Bademosi, Elizabeth Cooper-Williams, Isabel C. Morrow, Callista B. Harper, Woo Ram Jung, Robert G. Parton, Geoffrey J. Goodhill, Andreas Papadopulos, Frédéric A. Meunier

Research output: Contribution to journalArticlepeer-review

39 Scopus citations

Abstract

Our understanding of endocytic pathway dynamics is severely restricted by the diffraction limit of light microscopy. To address this, we implemented a novel technique based on the subdiffractional tracking of internalized molecules (sdTIM). This allowed us to image anti-green fluorescent protein Atto647N-tagged nanobodies trapped in synaptic vesicles (SVs) from live hippocampal nerve terminals expressing vesicle-associated membrane protein 2 (VAMP2)-pHluorin with 36-nm localization precision. Our results showed that, once internalized, VAMP2-pHluorin/Atto647N-tagged nanobodies exhibited a markedly lower mobility than on the plasma membrane, an effect that was reversed upon restimulation in presynapses but not in neighboring axons. Using Bayesian model selection applied to hidden Markov modeling, we found that SVs oscillated between diffusive states or a combination of diffusive and transport states with opposite directionality. Importantly, SVs exhibiting diffusive motion were relatively less likely to switch to the transport motion. These results highlight the potential of the sdTIM technique to provide new insights into the dynamics of endocytic pathways in a wide variety of cellular settings.

Original languageEnglish
Pages (from-to)277-292
Number of pages16
JournalJournal of Cell Biology
Volume215
Issue number2
DOIs
StatePublished - 2016

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