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Journal of Biomedical Optics

Combined lineage mapping and gene expression profiling of embryonic brain patterning using ultrashort pulse microscopy and image registration
Author(s): Holly C. Gibbs; Colin R. Dodson; Yuqiang Bai; Arne C. Lekven; Alvin T. Yeh
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Paper Abstract

During embryogenesis, presumptive brain compartments are patterned by dynamic networks of gene expression. The spatiotemporal dynamics of these networks, however, have not been characterized with sufficient resolution for us to understand the regulatory logic resulting in morphogenetic cellular behaviors that give the brain its shape. We have developed a new, integrated approach using ultrashort pulse microscopy [a high-resolution, two-photon fluorescence (2PF)-optical coherence microscopy (OCM) platform using 10-fs pulses] and image registration to study brain patterning and morphogenesis in zebrafish embryos. As a demonstration, we used time-lapse 2PF to capture midbrain-hindbrain boundary morphogenesis and a <italic<wnt1</italic< lineage map from embryos during brain segmentation. We then performed <italic<in situ</italic< hybridization to deposit NBT/BCIP, where <italic<wnt1</italic< remained actively expressed, and reimaged the embryos with combined 2PF-OCM. When we merged these datasets using morphological landmark registration, we found that the mechanism of boundary formation differs along the dorsoventral axis. Dorsally, boundary sharpening is dominated by changes in gene expression, while ventrally, sharpening may be accomplished by lineage sorting. We conclude that the integrated visualization of lineage reporter and gene expression domains simultaneously with brain morphology will be useful for understanding how changes in gene expression give rise to proper brain compartmentalization and structure.

Paper Details

Date Published: 24 December 2014
PDF: 14 pages
J. Biomed. Opt. 19(12) 126016 doi: 10.1117/1.JBO.19.12.126016
Published in: Journal of Biomedical Optics Volume 19, Issue 12
Show Author Affiliations
Holly C. Gibbs, Texas A&M Univ. (United States)
Colin R. Dodson, Texas A&M Univ. (United States)
Yuqiang Bai, Texas A&M Univ. (United States)
Arne C. Lekven, Texas A&M Univ. (United States)
Alvin T. Yeh, Texas A&M Univ. (United States)

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