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Quantitative PCR-based Assay to Measure Sonic Hedgehog Signaling in Cellular Model of Ciliogenesis
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Ptch2 mediates the Shh response in Ptch1-/- cells
Astrid C Alfaro1, Brock Roberts1, Lina Kwong1
1Department of Molecular and Cell Biology, 16 Barker Hall, 3204, University of California, Berkeley, CA 94720, USA.
Development (Cambridge, England)
|August 3, 2014
Summary
The patched 2 (Ptch2) protein suppresses sonic hedgehog (Shh) signaling during development. In the absence of Ptch1, Ptch2 mediates the Shh response, revealing its crucial role in pathway regulation.
Area of Science:
- Developmental Biology
- Molecular Biology
- Cell Signaling
Background:
- The sonic hedgehog (Shh) pathway is crucial for development and is regulated by interactions between the Shh ligand, patched 1 (Ptch1) receptor, and smoothened (Smo) activator.
- The prevailing model suggests Shh binding to Ptch1 releases Smo inhibition, activating the pathway.
Purpose of the Study:
- To investigate the role of patched 2 (Ptch2) in Shh signaling, particularly in the context of Ptch1 deficiency.
- To determine if Ptch2 functions as a Shh receptor or suppressor.
Main Methods:
- Analysis of Shh signaling in Ptch1 knockout cells and cells with dominant-negative Ptch1 and Ptch2 mutants in chick neural tube development.
- Utilizing Shh-blocking antibodies and assessing cell migration towards Shh.
Main Results:
- Ptch1(-/-) cells exhibit upregulated Shh response but remain ligand-dependent and Shh-sensitive.
- Dominant-negative Ptch2 expression activates Shh response, indicating a suppressive role.
- Ptch1(-/-);Ptch2(-/-) cells show no further Shh response activation, confirming Ptch2's role in mediating Shh response when Ptch1 is absent.
Conclusions:
- Patched 2 (Ptch2) acts as a suppressor of sonic hedgehog (Shh) signaling during early development.
- Ptch2 mediates Shh responses in cells lacking Ptch1, highlighting its functional significance in Shh pathway regulation.
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