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| Status |
Public on Nov 08, 2012 |
| Title |
β-catenin regulates FSHβ induction by GnRH: next generation RNA-Sequencing identifies Brms1L as a mediator of beta-catenin regulation of FSHβ gene expression |
| Organism |
Mus musculus |
| Experiment type |
Expression profiling by high throughput sequencing
|
| Summary |
The regulation of gonadotropin synthesis by GnRH (Gonadotropin-releasing hormone) plays an essential role in the neuroendocrine control of reproduction. The known signaling mechanisms involved in gonadotropin synthesis have been expanding. For example, involvement of β-catenin in LHβ induction by GnRH has been discovered. We examined the role of β-catenin in FSHβ gene expression in LβT2 gonadotrope cells. GnRH caused a sustained increase in nuclear β-catenin levels, which was significantly reduced by JNK inhibition. siRNA-mediated knockdown of β-catenin mRNA demonstrated that induction of FSHβ mRNA by GnRH depended on β-catenin and that regulation of FSHβ by β-catenin occurred independently of the JNK-c-jun pathway. β-catenin depletion had no impact on FSHβ mRNA stability. In LβT2 cells transfected with FSHβ promoter luciferase fusion constructs, GnRH responsiveness was conferred by the proximal promoter (-944/-1), and was markedly decreased by β-catenin knockdown. However, none of the TCF/LEF binding sites in that region were required for promoter activation by GnRH. Chromatin immunoprecipitation further corroborated the absence of direct interaction between β-catenin and the 1.8 kb FSHβ promoter. To elucidate the mechanism for the β-catenin effect, we analyzed ~1 billion reads of next generation RNA sequencing β-catenin knockdown assays and selected the nuclear cofactor Brms1L as one candidate for further study. Subsequent experiments confirmed that Brms1L mRNA expression was decreased by β-catenin knockdown as well as by JNK inhibition. Furthermore, knockdown of Brms1L significantly attenuated GnRH-induced FSHβ expression. Thus, our findings indicate that the expression of Brms1L depends on β-catenin activity and contributes to FSHβ induction by GnRH.
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| Overall design |
A total of 24 samples were analyzed, namely 6 different experimental conditions, each comprised of 4 replicates. Control samples are included in the analysis. For the RNA-Seq assay, samples were multiplexed in the form of 3 samples per lanes, resulting in a total of 8 lanes.
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| Contributor(s) |
Wang Q, Chikina M, Zaslavsky E, Pincas H, Sealfon SC |
| Citation(s) |
23211523, 24591653, 29487567 |
| Submission date |
Nov 07, 2012 |
| Last update date |
May 15, 2019 |
| Contact name |
Maria Chikina |
| E-mail(s) |
mchikina@gmail.com
|
| Organization name |
Mount Sinai School of Medicine
|
| Street address |
Annenberg 14-88 One Gustave Levy Pl
|
| City |
New York |
| State/province |
NY |
| ZIP/Postal code |
10029 |
| Country |
USA |
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| Platforms (1) |
| GPL13112 |
Illumina HiSeq 2000 (Mus musculus) |
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| Samples (24)
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| Relations |
| BioProject |
PRJNA179158 |
| SRA |
SRP017110 |
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