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| Status |
Public on Oct 26, 2020 |
| Title |
FGF9 Promotes Mouse Spermatogonial Stem Cell Proliferation Mediated by p38 MAPK Signaling [bulk RNA-seq] |
| Organism |
Mus musculus |
| Experiment type |
Expression profiling by high throughput sequencing
|
| Summary |
Objectives: Fibroblast growth factor 9 (FGF9) is expressed by somatic cells in the seminiferous tubules, yet little information exists about its role in regulating spermatogonial stem cells (SSCs).
Materials and Methods: Fgf9 overexpression lentivirus was injected into mouse testes, and PLZF immunostaining was performed to investigate the effect of FGF9 on spermatogonia in vivo. Effect of FGF9 on SSCs was detected by transplanting cultured germ cells into tubules of testes. RNA-seq of bulk RNA and single-cell was performed to explore FGF9 working mechanisms. SB203580 was used to disrupt p38 MAPK pathway. p38 MAPK protein expression was detected by western blot and qPCR was performed to determine different gene expression. Small interfering RNA (siRNA) was used to knock down Etv5 gene expression in germ cells.
Results: Overexpression of Fgf9 in vivo resulted in arrested spermatogenesis and accumulation of undifferentiated spermatogonia. Exposure of germ cell cultures to FGF9 resulted in larger numbers of SSCs over time. Inhibition of p38 MAPK phosphorylation negated the SSC growth advantage provided by FGF9. Etv5 and Bcl6b gene expression was enhanced by FGF9 treatment. Gene knockdown of Etv5 disrupted the growth effect of FGF9 in cultured SSCs along with downstream expression of Bcl6b.
Conclusions: Taken together, this data indicates that FGF9 is an important regulator of SSC proliferation, operating through p38 MAPK phosphorylation and upregulating Etv5 and Bcl6b in turn.
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| Overall design |
bulk RNA seq: 4 biological replicates of cultured germ cells (labelled s6, s7, s8 and s9) of each of the following treatments: control (0ng/ml FGF), 1ng/ml FGF9, 20ng/ml FGF9 and 1ng/ml FGF2)
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| Contributor(s) |
Yang F, Whelan EC, Guan X, Deng B, Wang S, Sun J, Avarbock MR, Wu X, Brinster RL |
| Citation(s) |
33107118 |
| |
| Submission date |
Oct 25, 2020 |
| Last update date |
Jul 10, 2024 |
| Contact name |
Eoin Christopher Whelan |
| E-mail(s) |
ewhelan@vet.upenn.edu
|
| Phone |
2158988805
|
| Organization name |
University of Pennsylvania School of Veterinary Medicine
|
| Department |
Biomedical Sciences
|
| Lab |
Sasaki
|
| Street address |
3800 Spruce St
|
| City |
Philadelphia |
| State/province |
PA |
| ZIP/Postal code |
19104 |
| Country |
USA |
| |
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| Platforms (1) |
| GPL19057 |
Illumina NextSeq 500 (Mus musculus) |
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| Samples (16)
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| This SubSeries is part of SuperSeries: |
| GSE160063 |
FGF9 Promotes Mouse Spermatogonial Stem Cell Proliferation Mediated by p38 MAPK Signaling |
|
| Relations |
| BioProject |
PRJNA671713 |
| SRA |
SRP288502 |
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