Real-time single-molecule imaging in zebrafish embryos uncovers non-canonical translation
Bellec, M.; Mattonet, K.; Morisaki, T.; Lay, M.; Liang, J.; Avinens, D.; Martinet, V.; Muriaux, D.; Stasevich, T. J.; Dufourt, J.; Stainier, D. Y. R.
Show abstract
Spatiotemporal control of protein synthesis is critical for many biological processes, yet understanding when and where proteins are produced in vivo remains technically challenging, especially in vertebrate models. Here, we present real-time visualization of translation of a dorsoventral morphogen in zebrafish embryos. Using ALFA_array labeling and lattice light-sheet microscopy, we tracked the translation dynamics of Bmp2b. Single-molecule analysis revealed low ribosome occupancy, indicating tightly regulated Bmp2b synthesis. Substituting bmp2b UTRs with viral UTRs increased both the number of mRNAs undergoing translation and the translation efficiency per mRNA. Additional experiments using viral and actb2 5UTRs uncovered a previously unrecognized cap-independent translation mechanism for bmp2b. Thus, using a newly developed platform to dissect translational regulation in vivo, we find that complex regulatory strategies govern morphogen production during early vertebrate development.
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