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Published on: April 25, 2022
MiR-145 regulates steroidogenesis in mouse primary granulosa cells through targeting Crkl
Shuo Wang1, Weicheng Tang1, Lanfang Ma2
1Department of Obstetrics and Gynecology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei, People's Republic of China.
Aims:
It has been demonstrated that miR-145 is expressed in primordial follicles and modulates the initiation of primordial follicle development. We aimed to explore the function of miR-145 in mouse granulosa cells (mGCs).
Materials And Methods:
The proliferation and differentiation of GCs were examined via MTT, EDU assay, QRT-PCR, ELISA and electron microscope analysis. The target of miR-145 was determined by bioinformatics analysis and luciferase reporter assay and the molecular mechanisms were examined via western blot and quantitative Real-Time RT-PCR.
Key Findings:
We proved that down-regulation of miR-145 could inhibit GCs proliferation and differentiation. In addition, we provided evidence that Crkl was the target gene of miR-145. The miR-145 antagomir caused an increase in Crkl expression and activation of the JNK/p38 MAPK pathway. Overexpression of Crkl with pEGFP-N1-Crkl vector inhibited GCs differentiation and progesterone synthesis as well as activation of the JNK/p38 MAPK pathway.
Significance:
Our study shows that miR-145 targets Crkl and through the JNK/p38 MAPK signaling pathway promotes the GCs proliferation, differentiation, and steroidogenesis. MiR-145 may play an important role in the ovarian physiology and pathology.
Insights
MicroRNA-145 (miR-145) promotes mouse granulosa cell proliferation and differentiation by targeting Crkl and activating the JNK/p38 MAPK pathway. MiR-145 is crucial for ovarian function.
Area of Science:
- Reproductive Biology
- Molecular Endocrinology
- Cell Signaling
Background:
- MicroRNA-145 (miR-145) is implicated in primordial follicle development.
- Its specific role in mouse granulosa cells (mGCs) requires elucidation.
Purpose of the Study:
- To investigate the function of miR-145 in mouse granulosa cells.
- To identify the molecular targets and signaling pathways regulated by miR-145.
Main Methods:
- Cell proliferation and differentiation assays (MTT, EDU, QRT-PCR, ELISA, electron microscopy).
- Target identification using bioinformatics and luciferase reporter assays.
- Molecular mechanism analysis via Western blot and qRT-PCR.
Main Results:
- Down-regulation of miR-145 inhibited mGC proliferation and differentiation.
- Crk-like protein (Crkl) was identified as a direct target of miR-145.
- miR-145 antagomir increased Crkl expression and activated JNK/p38 MAPK.
- Crkl overexpression inhibited mGC differentiation and progesterone synthesis.
Conclusions:
- MiR-145 promotes mGC proliferation, differentiation, and steroidogenesis by targeting Crkl and activating the JNK/p38 MAPK pathway.
- MiR-145 plays a significant role in ovarian physiology and pathology.
