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Quantitative PCR-based Assay to Measure Sonic Hedgehog Signaling in Cellular Model of Ciliogenesis
Published on: January 31, 2025
Desert Hedgehog mediates stem Leydig cell differentiation through Ptch2/Gli1/Sf1 signaling axis
Changle Zhao1, Yongxun Chen1, Lei Liu1
1Integrative Science Center of Germplasm Creation in Western China (CHONGQING) Science City, Key Laboratory of Freshwater Fish Reproduction and Development (Ministry of Education), Laboratory of Aquatic Science of Chongqing, School of Life Sciences, Southwest University, Chongqing, China.
Elife
|June 29, 2026
Summary
Desert Hedgehog (Dhh) signaling is crucial for Leydig cell development. This study identifies the Dhh/Patched 2 (Ptch2)/Glioma-associated oncogene homolog 1 (Gli1)/steroidogenic factor 1 (Sf1) axis as essential for stem Leydig cell differentiation and function.
Area of Science:
- Endocrinology
- Developmental Biology
- Molecular Biology
Background:
- Desert Hedgehog (Dhh) mutations lead to Leydig cell dysfunction and androgen insufficiency.
- The precise mechanisms of Dhh signaling in Leydig cell lineage commitment are not fully understood.
Purpose of the Study:
- To elucidate the signaling pathway regulating stem Leydig cell (SLC) differentiation in Nile tilapia.
- To identify the specific roles of Dhh, its receptor, transcriptional effectors, and downstream targets in Leydig cell development.
Main Methods:
- CRISPR/Cas9 gene knockout in Nile tilapia.
- Stem Leydig cell (SLC) transplantation.
- In vitro and in vivo genetic rescue experiments.
- Luciferase reporter assays.
Main Results:
- A critical Dhh/Patched 2 (Ptch2)/Glioma-associated oncogene homolog 1 (Gli1)/steroidogenic factor 1 (Sf1) signaling axis was identified.
- Dhh regulates SLC differentiation, not survival.
- Ptch2 acts as the functional inhibitory receptor for Dhh.
- Gli1 is the primary transcriptional effector, activating Sf1 expression.
- Sf1 is indispensable for SLC differentiation.
Conclusions:
- The study delineates the Dhh-Ptch2-Gli1-Sf1 axis, a fundamental pathway for Leydig cell development.
- This axis is essential for regulating stem Leydig cell differentiation and ensuring proper testicular function.
- Findings provide critical insights into the endocrine regulation of Leydig cell lineage commitment.
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