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MIR-203 SECRETED IN EXTRACELLULAR VESICLES MEDIATES THE COMMUNICATION BETWEEN NEURAL CREST AND PLACODE CELLS REQUIRED FOR TRIGEMINAL GANGLIA FORMATION
microRNA
CRISPR/Cas9
Intercellular communication
Extracellular vesicles
MicroRNAs
Comunicação Celular
Proteína 9 Associada à CRISPR
Gânglio Trigeminal
Author
Affilliation
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, by using chick embryos, 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. By employing cell-specific electroporations for either miR-203 sponging or genomic editing using CRISPR/Cas9, we elucidated that neural crest cells serve as the source, while placode cells serve as the site of action 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 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.
Keywords
Trigeminal ganglionmicroRNA
CRISPR/Cas9
Intercellular communication
Extracellular vesicles
DeCS
Vesículas extracelularesMicroRNAs
Comunicação Celular
Proteína 9 Associada à CRISPR
Gânglio Trigeminal
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