Robo2 regulates synaptic oxytocin content by affecting actin dynamics
The regulation of neuropeptide level at the site of release is essential for proper neurophysiological functions. We focused on a prominent neuropeptide, oxytocin (OXT) in the zebrafish as an in vivo model to visualize and quantify OXT content at the resolution of a single synapse. We found that OXT...
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| Format: | Article |
| Language: | English |
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eLife Sciences Publications Ltd
2019-06-01
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| Series: | eLife |
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| Online Access: | https://elifesciences.org/articles/45650 |
| _version_ | 1811252998215565312 |
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| author | Savani Anbalagan Janna Blechman Michael Gliksberg Ludmila Gordon Ron Rotkopf Tali Dadosh Eyal Shimoni Gil Levkowitz |
| author_facet | Savani Anbalagan Janna Blechman Michael Gliksberg Ludmila Gordon Ron Rotkopf Tali Dadosh Eyal Shimoni Gil Levkowitz |
| author_sort | Savani Anbalagan |
| collection | DOAJ |
| description | The regulation of neuropeptide level at the site of release is essential for proper neurophysiological functions. We focused on a prominent neuropeptide, oxytocin (OXT) in 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 enriched with polymerized actin. Perturbation of actin filaments by either cytochalasin-D or conditional Cofilin expression resulted in decreased synaptic OXT levels. Genetic loss of robo2 or slit3 displayed decreased synaptic OXT content and robo2 mutants displayed reduced mobility of the actin probe Lifeact-EGFP in OXT synapses. Using a novel transgenic reporter allowing real-time monitoring of OXT-loaded vesicles, we show 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 Slit3-Robo2-Cdc42, which controls local actin dynamics, with the maintenance of synaptic neuropeptide levels. |
| first_indexed | 2024-04-12T16:43:07Z |
| format | Article |
| id | doaj.art-36ecaea9355a47b5864283b2ceb34bd6 |
| institution | Directory Open Access Journal |
| issn | 2050-084X |
| language | English |
| last_indexed | 2024-04-12T16:43:07Z |
| publishDate | 2019-06-01 |
| publisher | eLife Sciences Publications Ltd |
| record_format | Article |
| series | eLife |
| spelling | doaj.art-36ecaea9355a47b5864283b2ceb34bd62022-12-22T03:24:40ZengeLife Sciences Publications LtdeLife2050-084X2019-06-01810.7554/eLife.45650Robo2 regulates synaptic oxytocin content by affecting actin dynamicsSavani Anbalagan0Janna Blechman1Michael Gliksberg2Ludmila Gordon3Ron Rotkopf4Tali Dadosh5Eyal Shimoni6Gil Levkowitz7https://orcid.org/0000-0002-3896-1881Department of Molecular Cell Biology, Weizmann Institute of Science, Rehovot, IsraelDepartment of Molecular Cell Biology, Weizmann Institute of Science, Rehovot, IsraelDepartment of Molecular Cell Biology, Weizmann Institute of Science, Rehovot, IsraelDepartment of Molecular Cell Biology, Weizmann Institute of Science, Rehovot, IsraelBioinformatics Unit, LSCF, Weizmann Institute of Science, Rehovot, Israel; Electron Microscopy Unit, Weizmann Institute of Science, Rehovot, IsraelDepartment of Chemical Research Support, Weizmann Institute of Science, Rehovot, IsraelDepartment of Chemical Research Support, Weizmann Institute of Science, Rehovot, IsraelDepartment of Molecular Cell Biology, Weizmann Institute of Science, Rehovot, IsraelThe regulation of neuropeptide level at the site of release is essential for proper neurophysiological functions. We focused on a prominent neuropeptide, oxytocin (OXT) in 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 enriched with polymerized actin. Perturbation of actin filaments by either cytochalasin-D or conditional Cofilin expression resulted in decreased synaptic OXT levels. Genetic loss of robo2 or slit3 displayed decreased synaptic OXT content and robo2 mutants displayed reduced mobility of the actin probe Lifeact-EGFP in OXT synapses. Using a novel transgenic reporter allowing real-time monitoring of OXT-loaded vesicles, we show 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 Slit3-Robo2-Cdc42, which controls local actin dynamics, with the maintenance of synaptic neuropeptide levels.https://elifesciences.org/articles/45650oxytocinarginine-vasopressinhypothalamusneuroendocrineneurohypophysissynapse |
| spellingShingle | Savani Anbalagan Janna Blechman Michael Gliksberg Ludmila Gordon Ron Rotkopf Tali Dadosh Eyal Shimoni Gil Levkowitz Robo2 regulates synaptic oxytocin content by affecting actin dynamics eLife oxytocin arginine-vasopressin hypothalamus neuroendocrine neurohypophysis synapse |
| title | Robo2 regulates synaptic oxytocin content by affecting actin dynamics |
| title_full | Robo2 regulates synaptic oxytocin content by affecting actin dynamics |
| title_fullStr | Robo2 regulates synaptic oxytocin content by affecting actin dynamics |
| title_full_unstemmed | Robo2 regulates synaptic oxytocin content by affecting actin dynamics |
| title_short | Robo2 regulates synaptic oxytocin content by affecting actin dynamics |
| title_sort | robo2 regulates synaptic oxytocin content by affecting actin dynamics |
| topic | oxytocin arginine-vasopressin hypothalamus neuroendocrine neurohypophysis synapse |
| url | https://elifesciences.org/articles/45650 |
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