Bernardi, Yanel E. and Sanchez-Vasquez, Estefania and Piacentino, Michael L. and Urrutia, Hugo and Rossi, Izadora and Alcantara Saraiva, Karina L. and Pereira-Neves, Antonio and Ramirez, Marcel Ivan and Bronner, Marianne E. and de Miguel, Natalia and Strobl-Mazzulla, Pablo Hernan (2023) Extracellular vesicle-localized miR-203 mediates neural crest-placode communication required for trigeminal ganglia formation. . (Unpublished) https://resolver.caltech.edu/CaltechAUTHORS:20230316-181901000.3
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Abstract
While interactions between neural crest and placode cells are critical for the proper formation of the trigeminal ganglion, the mechanisms underlying this process remain largely uncharacterized. Here, we show that the microRNA-(miR)203, whose epigenetic repression is required for neural crest migration, is reactivated in coalescing and condensing trigeminal ganglion cells. Overexpression of miR-203 induces ectopic coalescence of neural crest cells and increases ganglion size. Reciprocally, loss of miR-203 function in placode, but not neural crest, cells perturbs trigeminal ganglion condensation. Demonstrating intercellular communication, overexpression of miR-203 in the neural crest in vitro or in vivo represses a miR-responsive sensor in placode cells. Moreover, neural crest-secreted extracellular vesicles (EVs), visualized using pHluorin-CD63 vector, become incorporated into the cytoplasm of placode cells. Finally, RT-PCR analysis shows that small EVs isolated from condensing trigeminal ganglia are selectively loaded with miR-203. Together, our findings reveal a critical role in vivo for neural crest-placode communication mediated by sEVs and their selective microRNA cargo for proper trigeminal ganglion formation.
Item Type: | Report or Paper (Discussion Paper) | ||||||||||||||||||
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Additional Information: | 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. We thank all the authors and members in the Laboratory of Developmental Biology at the INTECH for their contribution and helpful discussions during the course of our study. We thank Dr. Alissa M. Weaver (Vanderbilt University School of Medicine, Nashville, TN, USA) for the pHluo construct. We are very grateful to the directors and students from “Escuela de Educación Secundaria Agraria de Chascomús” for providing fertilized eggs of excellent quality. This work was supported by the Agencia Nacional de Promoción Cientı́fica y Tecnológica (PICT 2018-1879 to P.H.S-M.) and by the Fogarty International Center of the National Institutes of Health (R21TW011224 to M.E.B. and P.H.S-M.) Author contributions: Y.E.B. and P.H.S-M. designed, performed the experiments and wrote the manuscript with editing and input from all coauthors; E.S-V performed experiments; M.L.P. and H.U. contributed in the co-culture and time-lapse experiments; I.R and M.I.R contributed on the NTA analysis of sEVs; K.L.A.S and A.P-N. performed EM imaging and aided in their interpretation; M.E.B contributed in the discussion, experimental design and manuscript writing; N.d.M. contributed on the sEVs purifications, characterization, experimental design and manuscript writing. The authors have declared no competing interest. | ||||||||||||||||||
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DOI: | 10.1101/2023.03.14.532527 | ||||||||||||||||||
Record Number: | CaltechAUTHORS:20230316-181901000.3 | ||||||||||||||||||
Persistent URL: | https://resolver.caltech.edu/CaltechAUTHORS:20230316-181901000.3 | ||||||||||||||||||
Usage Policy: | No commercial reproduction, distribution, display or performance rights in this work are provided. | ||||||||||||||||||
ID Code: | 120122 | ||||||||||||||||||
Collection: | CaltechAUTHORS | ||||||||||||||||||
Deposited By: | George Porter | ||||||||||||||||||
Deposited On: | 22 Mar 2023 17:04 | ||||||||||||||||||
Last Modified: | 22 Mar 2023 17:04 |
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