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

Joensuu, Merja, Padmanabhan, Pranesh, Durisic, Nela, Bademosi, Adekunle T. D., Cooper-Williams, Elizabeth, Morrow, Isabel C., Harper, Callista B., Jung, WooRam, Parton, Robert G., Goodhill, Geoffrey J., Papadopulos, Andreas and Meunier, Frederic A. (2016) Subdiffractional tracking of internalized molecules reveals heterogeneous motion states of synaptic vesicles. Journal of Cell Biology, 215 2: 277-292. doi:10.1083/jcb.201604001


Author Joensuu, Merja
Padmanabhan, Pranesh
Durisic, Nela
Bademosi, Adekunle T. D.
Cooper-Williams, Elizabeth
Morrow, Isabel C.
Harper, Callista B.
Jung, WooRam
Parton, Robert G.
Goodhill, Geoffrey J.
Papadopulos, Andreas
Meunier, Frederic A.
Title Subdiffractional tracking of internalized molecules reveals heterogeneous motion states of synaptic vesicles
Journal name Journal of Cell Biology   Check publisher's open access policy
ISSN 1540-8140
0021-9525
Publication date 2016-10-24
Year available 2016
Sub-type Article (original research)
DOI 10.1083/jcb.201604001
Open Access Status DOI
Volume 215
Issue 2
Start page 277
End page 292
Total pages 16
Place of publication New York, NY, United States
Publisher Rockefeller University Press
Language eng
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.
Keyword Cell Biology
Cell Biology
Q-Index Code C1
Q-Index Status Provisional Code
Grant ID DP120104057
1037320
Institutional Status UQ

 
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