Spatiotemporal Characterization of Anterior Segment Mesenchyme Heterogeneity During Zebrafish Ocular Anterior Segment Development

Frontiers in Cell and Developmental Biology
Kristyn L Van Der MeulenJakub K Famulski

Abstract

Assembly of the ocular anterior segment (AS) is a critical event during development of the vertebrate visual system. Failure in this process leads to anterior segment dysgenesis (ASD), which is characterized by congenital blindness and predisposition to glaucoma. The anterior segment is largely formed via a neural crest-derived population, the Periocular Mesenchyme (POM). In this study, we aimed to characterize POM behaviors and transcriptional identities during early establishment of the zebrafish AS. Two-color fluorescent in situ hybridization suggested that early AS associated POM comprise of a heterogenous population. In vivo and time-course imaging analysis of POM distribution and migratory dynamics analyzed using transgenic zebrafish embryos (Tg[foxc1b:GFP], Tg[foxd3:GFP], Tg[pitx2:GFP], Tg[lmx1b.1:GFP], and Tg[sox10:GFP]) revealed unique AS distribution and migratory behavior among the reporter lines. Based on fixed timepoint and real-time analysis of POM cell behavior a comprehensive model for colonization of the zebrafish AS was assembled. Furthermore, we generated single cell transcriptomic profiles (scRNA) from our POM reporter lines and characterized unique subpopulation expression patterns. Based on scRNA clusterin...Continue Reading

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Citations

Jan 12, 2021·Frontiers in Cell and Developmental Biology·Antionette L Williams, Brenda L Bohnsack

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Methods Mentioned

BETA
transgenic
scRNA assay
PCR
Flow Cytometry
chip

Software Mentioned

Nikon Elements Viewer
GraphPad Prism8
Adobe Photoshop
Adobe Illustrator
Cell Ranger3
FWISH
Elements
Loupe
Nikon Elements
Excel

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