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Three-dimensional morphology and gene expression in the Drosophila blastoderm at cellular resolution I: data acquisition pipeline

Journal Article · · GenomeBiology.com
 [1];  [2];  [3];  [2];  [4];  [2];  [2];  [2];  [4];  [2];  [3];  [2];  [2];  [2]
  1. Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States); DOE/OSTI
  2. Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)
  3. University of California, Berkeley, CA (United States)
  4. University of California, Davis, CA (United States)

To model and thoroughly understand animal transcription networks, it is essential to derive accurate spatial and temporal descriptions of developing gene expression patterns with cellular resolution. Here we describe a suite of methods that provide the first quantitative three-dimensional description of gene expression and morphology at cellular resolution in whole embryos. A database containing information derived from 1,282 embryos is released that describes the mRNA expression of 22 genes at multiple time points in the Drosophila blastoderm. We demonstrate that our methods are sufficiently accurate to detect previously undescribed features of morphology and gene expression. The cellular blastoderm is shown to have an intricate morphology of nuclear density patterns and apical/basal displacements that correlate with later well-known morphological features. Pair rule gene expression stripes, generally considered to specify patterning only along the anterior/posterior body axis, are shown to have complex changes in stripe location, stripe curvature, and expression level along the dorsal/ventral axis. Pair rule genes are also found to not always maintain the same register to each other. The application of these quantitative methods to other developmental systems will likely reveal many other previously unknown features and provide a more rigorous understanding of developmental regulatory networks.

Research Organization:
Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)
Sponsoring Organization:
USDOE Office of Science (SC), Biological and Environmental Research (BER); Academy of Finland; National Institute of General Medical Sciences (NIGMS); National Human Genome Research Institute (NHGRI)
Grant/Contract Number:
AC02-05CH11231
OSTI ID:
1626708
Journal Information:
GenomeBiology.com, Journal Name: GenomeBiology.com Journal Issue: 12 Vol. 7; ISSN 1465-6906
Publisher:
BioMed CentralCopyright Statement
Country of Publication:
United States
Language:
English

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High-speed panoramic light-sheet microscopy reveals global endodermal cell dynamics journal July 2013
A microfluidic array for large-scale ordering and orientation of embryos journal December 2010
The appeasement of Doug: a synthetic approach to enhancer biology journal March 2016
Quantitative imaging of the Dorsal nuclear gradient reveals limitations to threshold-dependent patterning in Drosophila journal December 2009
Multimodal transcriptional control of pattern formation in embryonic development journal December 2019
Bioimage informatics: a new area of engineering biology journal July 2008
Simultaneous recognition and segmentation of cells: application in C.elegans journal August 2011
Automated cellular annotation for high-resolution images of adult Caenorhabditis elegans journal June 2013
Fly-on-a-Chip: Microfluidics for Drosophila melanogaster Studies journal December 2019
Multimodal transcriptional control of pattern formation in embryonic development posted_content January 2019
Quantitative comparison of the anterior-posterior patterning system in the embryos of five Drosophila species journal January 2018
Dissecting the sharp response of a canonical developmental enhancer reveals multiple sources of cooperativity journal January 2018
An in silico analysis of robust but fragile gene regulation links enhancer length to robustness posted_content June 2019
Single-cell resolution view of the transcriptional landscape of developing Drosophila eye posted_content January 2019
Multicolor fluorescent in situ hybridization to define abutting and overlapping gene expression in the embryonic zebrafish brain journal January 2011
VIPER: Visualization Pipeline for RNA-seq, a Snakemake workflow for efficient and complete RNA-seq analysis journal April 2018
Synthetic enhancer design by in silico compensatory evolution reveals flexibility and constraint in cis-regulation journal November 2017
An in silico analysis of robust but fragile gene regulation links enhancer length to robustness journal November 2019
Hunchback is counter-repressed to regulate even-skipped stripe 2 expression in Drosophila embryos journal September 2018
Quantitative Comparison of the Anterior-Posterior Patterning System in the Embryos of Five Drosophila Species journal July 2019
Depleting Gene Activities in Early Drosophila Embryos with the “Maternal-Gal4–shRNA” System journal January 2013
Mathematical modeling of gene expression: a guide for the perplexed biologist journal January 2011
Dissecting the sharp response of a canonical developmental enhancer reveals multiple sources of cooperativity journal June 2019

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