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Robo2 regulates synaptic oxytocin content by affecting actin state

View ORCID ProfileSavani Anbalagan, Janna Blechman, Michael Gliksberg, Ron Rotkopf, Tali Dadosh, View ORCID ProfileGil Levkowitz
doi: https://doi.org/10.1101/553198
Savani Anbalagan
1Department of Molecular Cell Biology, Weizmann Institute of Science, PO Box 26, Rehovot 7610001, Israel
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  • ORCID record for Savani Anbalagan
Janna Blechman
1Department of Molecular Cell Biology, Weizmann Institute of Science, PO Box 26, Rehovot 7610001, Israel
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Michael Gliksberg
1Department of Molecular Cell Biology, Weizmann Institute of Science, PO Box 26, Rehovot 7610001, Israel
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Ron Rotkopf
2Bioinformatics Unit, LSCF, Weizmann Institute of Science, PO Box 26, Rehovot 7610001, Israel
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Tali Dadosh
3Electron Microscopy Unit, Department of Chemical Research Support, Weizmann Institute of Science, PO Box 26, Rehovot 7610001, Israel
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Gil Levkowitz
1Department of Molecular Cell Biology, Weizmann Institute of Science, PO Box 26, Rehovot 7610001, Israel
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  • ORCID record for Gil Levkowitz
  • For correspondence: gil.levkowitz@weizmann.ac.il
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Summary

The regulation of neuropeptide level at the site of release is essential for proper neurophysiological functions. We focused on a prominent neuropeptide, oxytocin (OXT), and used the zebrafish as an in vivo model to visualize and quantify OXT content at the resolution of a single synapse. We found that OXT-loaded synapses were surrounded by polymerized actin. Perturbation of actin filaments by cytochalasin-D resulted in decreased synaptic OXT levels. Live imaging of the actin probe, Lifeact-EGFP, showed reduced mobility in OXT synapses in robo2 mutants, which displayed decreased synaptic OXT content. Using a novel transgenic reporter line allowing real-time monitoring of OXT-loaded vesicles, we showed that robo2 mutants display slower rate of vesicles accumulation. OXT-specific expression of dominant-negative Cdc42, which is a key regulator of actin dynamics and a downstream effector of Robo2, led to a dose-dependent increase in OXT content in WT, and a dampened effect in robo2 mutants. Our results link Robo2-Cdc42 signalling, which controls local actin dynamics, with the maintenance of synaptic neuropeptide levels.

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The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY 4.0 International license.
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Posted February 17, 2019.
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Robo2 regulates synaptic oxytocin content by affecting actin state
Savani Anbalagan, Janna Blechman, Michael Gliksberg, Ron Rotkopf, Tali Dadosh, Gil Levkowitz
bioRxiv 553198; doi: https://doi.org/10.1101/553198
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Robo2 regulates synaptic oxytocin content by affecting actin state
Savani Anbalagan, Janna Blechman, Michael Gliksberg, Ron Rotkopf, Tali Dadosh, Gil Levkowitz
bioRxiv 553198; doi: https://doi.org/10.1101/553198

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