miR-203 secreted in extracellular vesicles mediates the communication between neural crest and placode cells required for trigeminal ganglia formation

BERNARDI, YANEL E.; SANCHEZ-VASQUEZ, ESTEFANIA; MÁRQUEZ, ROCÍO BELÉN; PIACENTINO, MICHAEL L.; URRUTIA, HUGO; ROSSI, IZADORA; ALCÂNTARA SARAIVA, KARINA L.; PEREIRA-NEVES, ANTONIO; RAMIREZ, MARCEL I.; BRONNER, MARIANNE E.; DE MIGUEL, NATALIA; STROBL-MAZZULLA, PABLO H.

PLOS Biology

PLOS

Año: 2024 vol. 22

AWUhi:lePilnetaesreaccotniofinrms tbheattwalelheenandeinugrlaelvcelrseasrtearenpdrepsleanctoeddceocrerellcstlayr:e critical for the proper formationof the trigeminal ganglion, the mechanisms underlying this process remain largelyuncharacterized. Here, by using chick embryos, we show that the microRNA (miR)-203,whose epigenetic repression is required for neural crest migration, is reactivated in coalescingand condensing trigeminal ganglion cells. Overexpression of miR-203 induces ectopiccoalescence of neural crest cells and increases ganglion size. By employing cell-specificelectroporations for either miR-203 sponging or genomic editing using CRISPR/Cas9, weelucidated that neural crest cells serve as the source, while placode cells serve as the site ofaction for miR-203 in trigeminal ganglion condensation. Demonstrating intercellular communication,overexpression of miR-203 in the neural crest in vitro or in vivo represses an miRresponsivesensor in placode cells. Moreover, neural crest-secreted extracellular vesicles(EVs), visualized using pHluorin-CD63 vector, become incorporated into the cytoplasm ofplacode cells. Finally, RT-PCR analysis shows that small EVs isolated from condensing trigeminalganglia are selectively loaded with miR-203. Together, our findings reveal a criticalrole in vivo for neural crest-placode communication mediated by sEVs and their selectivemicroRNA cargo for proper trigeminal ganglion formation.