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Expression profiling of Noto-GFP+ notochord progenitor cells sorted from E8.5 mouse embryos

ABSTRACT: Using a combination of cell sorting and microarray analysis, we identified almost 200 genes as having a high level of expression in the notochord. After whole mount in situ hybridization screening, we confirmed approximately one third of these as having a novel notochord expression pattern. Keywords: cell type comparison - embryonic Noto-GFP+ notochord progenitors versus surrounding GFP- comparator cells

ORGANISM(S): Mus musculus

PROVIDER: GSE14211 | GEO | 2011/10/01

SECONDARY ACCESSION(S): PRJNA111337

REPOSITORIES: GEO

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Expression profiling of Noto-GFP+ notochord progenitor cells sorted from E8.5 mouse embryos

Project description:Using a combination of cell sorting and microarray analysis, we identified almost 200 genes as having a high level of expression in the notochord. After whole mount in situ hybridization screening, we confirmed approximately one third of these as having a novel notochord expression pattern. Keywords: cell type comparison - embryonic Noto-GFP+ notochord progenitors versus surrounding GFP- comparator cells 3 biological replicates - each replicate includes an experiment cell population (Noto-GFP positive cell sort) and a comparator population (GFP negative cell sort)

2011-09-30 | E-GEOD-14211 | biostudies-arrayexpress

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Project description:We screened for differentially expressed genes in the developing notochord using the Affymetrix microarray system in Xenopus laevis. At late gastrula, we dissected four regions from the embryo, anterior mesoderm, posterior mesoderm, notochord and presomitic mesoderm. Three types of comparison were carried out to generate a list of predominantly notochord expressed genes: (1) Posterior mesoderm vs. anterior mesoderm; notochord genes are expected to be increased since the notochord is located in the posterior mesoderm. (2) Posterior mesoderm vs. whole embryos; notochord genes are expected to be increased. (3) Notochord vs. somite. This comparison sub-divided the group of posterior mesodermal genes identified in (1) and (2). All tissues are dissected using tungsten needles. We first dissected dorsal tissue above the archenteron from late gastrula to early neurula. To loosen tissue, we treated the dissected dorsal explant in a 1% cysteine solution (pH 7.4) and removed the neuroectodermal layer. Anterior mesoderm was dissected corresponding to about the anterior one-third of the archenteron roof, and the rest was collected as posterior mesoderm. The posterior mesodermal explant was dissected into notochord and somites, following a clearly visible border between the two tissues. The accuracy of all dissection was confirmed by RT-PCR of marker genes.

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Project description:Expression profiling of Noto-GFP+ notochord progenitor cells sorted from E8.5 mouse embryos

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