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| Status |
Public on May 01, 2007 |
| Title |
Identif. of oscillatory genes in somitogenesis from functional genomic analysis of a human mesenchymal stem cell model |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by array
|
| Summary |
During somitogenesis, oscillatory expression of genes in the notch and wnt signaling pathways plays a key role in regulating segmentation. These oscillations in expression levels are elements of a species-specific developmental mechanism. To date, the periodicity and components of the human clock remain unstudied. Here we show that a human mesenchymal stem/stromal cell (MSC) model can be induced to display oscillatory gene expression. We observed that the known cycling gene HES1 oscillated with a 5 hour period, consistent with available data on the rate of somitogenesis in humans. We also observed cycling of Hes1 expression in mouse C2C12 myoblasts with a period of 2 hours, consistent with previous in vitro and embryonic studies. Furthermore, we used microarray and quantitative PCR (Q-PCR) analysis to identify additional genes that display oscillatory expression both in vitro and in mouse embryos. We confirmed oscillatory expression of the notch pathway gene Maml3 and the wnt pathway gene Nkd2 by whole mount in situ hybridization analysis and Q-PCR. Expression patterns of these genes were disrupted in Wnt3atm1Amc mutants but not in Dll3pu mutants. Our results demonstrate that human and mouse in vitro models can recapitulate oscillatory expression observed in embryo and that a number of genes in multiple developmental pathways display dynamic expression in vitro.
Keywords: time series
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| Overall design |
Synchronization of human umbilical cord blood derived stem/stromal cell (population 1) was carried out as follows: T-25 cm flasks were set up in parallel. These cells were grown to 90% confluence in DMEM supplemented with 10% FBS (UCB-MSC), then incubated in DMEM with only 0.2% FBS for 24 hours, and returned to DMEM supplemented with FBS. Samples were collected at 30 min. intervals from 0 to 8 hours, and then at 1 hour intervals from 9 to 24 hours.
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| Contributor(s) |
William DA, Saitta B, Gibson JD, Traas J, Markov V, Gonzalez DM, Sewell W, Anderson DM, Pratt SC, Rappaport EF, Kusumi K |
| Citation(s) |
17362910 |
| |
| Submission date |
Feb 12, 2007 |
| Last update date |
Aug 10, 2018 |
| Contact name |
Kenro Kusumi |
| E-mail(s) |
[email protected]
|
| Phone |
480-727-8993
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| Fax |
480-965-6899
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| URL |
http://sols.asu.edu/faculty/kkusumi.php
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| Organization name |
Arizona State University
|
| Department |
School of Life Sciences
|
| Street address |
PO Box 874501
|
| City |
Tempe |
| State/province |
AZ |
| ZIP/Postal code |
85287 |
| Country |
USA |
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| Platforms (2) |
| GPL96 |
[HG-U133A] Affymetrix Human Genome U133A Array |
| GPL97 |
[HG-U133B] Affymetrix Human Genome U133B Array |
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| Samples (38)
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| This SubSeries is part of SuperSeries: |
| GSE7054 |
Identification of oscillatory genes in somitogenesis from functional genomic analysis |
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| Relations |
| BioProject |
PRJNA104045 |
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