The content and activity of extracellular vesicles purified from cell culture media or bodily fluids have been studied extensively; however, the physiological relevance of exosomes within normal biological systems is poorly characterized, particularly during development. Although exosomes released by invasive, metastatic cells alter migration of neighboring cells in culture, it is unclear whether cancer cells misappropriate exosomes released by healthy, differentiated cells or reactivate dormant developmental programs that include exosome cell-cell communication. Using chick cranial neural fold cultures, we show that migratory neural crest cells, a developmentally critical cell type and model for metastasis, release and deposit CD63-positive, 30-100 nm particles into the extracellular environment. Neural crest cells contain ceramide-rich multi-vesicular bodies and produce larger vesicles positive for migrasome markers as well. We conclude that neural crest cells produce extracellular vesicles including exosomes and migrasomes. When Rab27a plasma membrane docking is inhibited, neural crest cells become less polarized and rounded, leading to a loss of directional migration and reduced speed. These results indicate that neural crest cell exosome release is critical for migration.
Bibliographical noteFunding Information:
under Award Number ECCS-1542202. Imaging was also supported by the resources and staff
We are grateful to the Verweij lab for the kind gift of the CD63-pHluorin construct. We thank Yasuhiko Kawakami, Meg Titus, and members of the Gammill and Kawakami labs for discussion and advice. Thanks also to Aidan Peterson and the Titus lab for assistance with confocal imaging, and to Micah Gearhart for assistance with RNAseq data analysis. Portions of this work were conducted in the Minnesota Nano Center, which is supported by the National Science Foundation through the National Nano Coordinated Infrastructure Network (NNCI) under Award Number ECCS-1542202. Imaging was also supported by the resources and staff at the University of Minnesota University Imaging Centers (UIC). SCR_020997. Special thanks to Patrick Willey. The monoclonal antibody, HNK-1 (clone 3H5) was developed by W.M. Halfter from the University of Pittsburgh, and was obtained from the Developmental Studies Hybridoma Bank, created by the NICHD of the NIH and maintained at The University of Iowa, Department of Biology, Iowa City, IA 52242 This work was supported by: a University of Minnesota Grant-in-Aid [to L.S.G.]; National Science Foundation Graduate Research Fellowship [Project No. 00074041 to C.M.G]; and the National Institutes of Health [T90 DE022732 to C.M.G. via the Minnesota Craniofacial Research Training Program]. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institute of Dental & Craniofacial Research or the National Institutes of Health.
Science Foundation through the National Nano Coordinated Infrastructure Network (NNCI)
© 2022. Published by The Company of Biologists Ltd.
- extracellular vesicles
- neural crest
PubMed: MeSH publication types
- Journal Article
- Research Support, Non-U.S. Gov't
- Research Support, N.I.H., Extramural