It's been a fantastic experience to publish our work in @natcomms.nature.com and I sincerely thank the reviewers of our paper for their constructive and helpful comments! (8/8)

I deeply thank all co-authors for their constant support over the past years: @uljakravcenko.bsky.social, @mohsensadeghi.bsky.social, Christoph Diebolder, Malgorzata Lubas, Lia Ivanov, Thiemo Sprink, Magdalena Schacherl, @mishakudryashev.bsky.social and Christian Rosenmund. (7/8)

Intriguingly, we observed filamentous connectors between the majority of fusing vesicles and at least one more adjacent vesicle. We suggest that these interconnectors physically couple vesicle fusion and direct vesicle resupply required for release site refilling. (6/8)

Our morphometric and computational analyses revealed that synaptic vesicle fusion is likely initiated via stalk formation, whereby vesicle and cell membrane form a tip-like connection. We did not find evidence for tight docking or strong membrane bending prior to fusion. (5/8)

In these stimulated synapses, we identified more than 50 synaptic vesicle fusion intermediates and characterized them morphometrically at nanoscale resolution and under near-native conditions. (4/8)

We confirmed successful optogenetic stimulation of synapses directly via cryo-confocal microscopy of the glutamate sensor iGluSnFR3. Neurotransmitters, released during exocytosis, induced an increase in fluorescence intensity that was preserved under cryo conditions. (3/8)

We established a timed in situ cryo-electron tomography workflow to capture the complete sequence of synaptic vesicle fusion in optogenetically stimulated mouse neurons. Using a modified plunge freezer, the "Voptobot", we stimulated neurons only 2-3 milliseconds before cryofixation. (2/8)

Thank you so much! 😊

In the left image, I see a queen with a crown🫅 But I guess that's not what you meant? 😅