PUBLICATION
Combined lineage mapping and gene expression profiling of embryonic brain patterning using ultrashort pulse microscopy and image registration
- Authors
- Gibbs, H.C., Dodson, C.R., Bai, Y., Lekven, A.C., Yeh, A.T.
- ID
- ZDB-PUB-150219-10
- Date
- 2014
- Source
- Journal of Biomedical Optics 19: 126016 (Journal)
- Registered Authors
- Lekven, Arne
- Keywords
- none
- MeSH Terms
-
- Animals
- Brain Chemistry/physiology
- In Situ Hybridization
- Gene Expression Profiling/methods*
- Microscopy, Fluorescence, Multiphoton/methods*
- Brain/embryology*
- Brain/metabolism
- Zebrafish
- Wnt Signaling Pathway
- Imaging, Three-Dimensional/methods*
- Embryo, Nonmammalian
- Tomography, Optical Coherence/methods*
- PubMed
- 25539062 Full text @ J. Biomed. Opt.
Citation
Gibbs, H.C., Dodson, C.R., Bai, Y., Lekven, A.C., Yeh, A.T. (2014) Combined lineage mapping and gene expression profiling of embryonic brain patterning using ultrashort pulse microscopy and image registration. Journal of Biomedical Optics. 19:126016.
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 wnt1 lineage map from embryos during brain segmentation. We then performed in situ hybridization to deposit NBT/BCIP, where wnt1 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.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping