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| Status |
Public on Feb 24, 2020 |
| Title |
Differentially expressed transcriptomes of P7 mouse tendon cells with targeted deletion of TGF-beta signaling |
| Organism |
Mus musculus |
| Experiment type |
Expression profiling by high throughput sequencing
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| Summary |
Background: Our group has previously shown that disruption of TGFβ signaling in mouse limb mesenchyme resulted in arrested tendon formation (Pryce et at, 2007). To examine the role of TGFβ signaling in later stages of tendon development, the TGF-beta type II receptor gene (Tgfbr2) was targeted in the Scleraxis (Scx)-expressing cell lineage using the Cre-lox recombination system. We find that tendon development was not disrupted in mutant (Tgfbr2;ScxCre) embryos. However, shortly after birth tenocytes underwent dedifferentiation in which the cell lost differentiation markers and reverted to a more stem/progenitor state.
Purpose: To determine gene expression changes in Tgfbr2;ScxCre mutant tendon cells.
Methods: We performed scRNA-seq for transcriptome changes in P7 mutant tendon cells, a stage at which the majority of the mutant cells is dedifferentiated. Briefly, tendons from P7 mutant and wild-type (as a control) pups were harvested and enzymatically digested. The released cells were then subjected to scRNA-seq analysis using 10x Genomics platform.
Results: Using unsupervised hierarchical clustering, we identified two major clusters corresponding to mutant (dedifferentiated) cells and wild-type tenocytes in the respective samples. Findings from the pairwise comparison of the gene set between the P7 wild-type tenocyte and mutant cell clusters do not only lend support to our notion that the mutant cells lost their differentiation state, but also suggest the possibility of induction of some developmental programs in these cells, a general feature in cellular dedifferentiation.
Conclusions: TGF-beta signaling is critical for maintenance of the tendon cell fate.
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| Overall design |
Tendon mRNA profiles of 7-day old Tgfbr2;ScxCre mutant and wild type (as a control) mice were generated by deep sequencing using Illumina NextSeq (MidOutput 150). Two samples were submitted, in which each sample consist of enzymatically-released tendon cells that were harvested and pooled from 2 pups.
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| Contributor(s) |
Tan G, Wang C, Xia Z, Schweitzer R |
| Citation(s) |
31961320 |
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| Submission date |
Oct 29, 2019 |
| Last update date |
Feb 24, 2020 |
| Contact name |
Zheng Xia |
| E-mail(s) |
xiaz@ohsu.edu
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| Organization name |
Oregon Health & Science University
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| Department |
Department of Biomedical Engineering
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| Lab |
Xia Lab
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| Street address |
3181 SW Sam Jackson Park Rd
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| City |
Portland |
| State/province |
Oregon |
| ZIP/Postal code |
97239 |
| Country |
USA |
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| Platforms (1) |
| GPL19057 |
Illumina NextSeq 500 (Mus musculus) |
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| Samples (2) |
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| Relations |
| BioProject |
PRJNA580249 |
| SRA |
SRP227361 |
