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Updated: Apr 29, 2026
Modeling Paracrine Noncanonical Wnt Signaling In Vitro
Published on: December 10, 2021
Concentration-dependent effects of WNTLESS on WNT1/3A signaling
Lisa M Galli1, Linda A Szabo, Lydia Li
1Department of Biology, San Francisco State University, San Francisco, California.
Background:
WNTLESS (WLS) is a multi-transmembrane protein that transports Wnt ligands from the Golgi to the cell surface. Although WLS loss-of-function experiments in the developing central nervous system reveal phenotypes consistent with defects in WNT1 and WNT3A signaling, data from complementary gain-of-function experiments have not yet been reported. Here, we report the phenotypic consequences of WLS overexpression in cultured cells and in the developing chick spinal cord.
Results:
Overexpression of small amounts of WLS along with either WNT1 or WNT3A promotes the Wnt/β-catenin pathway in HEK293T cells, while overexpression of higher levels of WLS inhibits the Wnt/β-catenin pathway in these cells. Similarly, overexpressed WLS inhibits the Wnt/β-catenin pathway in the developing spinal cord, as assessed by cell proliferation and specification. These effects appear to be Wnt-specific as overexpression of WLS inhibits the expression of FZD10, a target of β-catenin-dependent transcription.
Conclusions:
Our results show that overexpression of WLS inhibits Wnt/β-catenin signaling in the spinal cord. As the activation of the Wnt/β-catenin pathway in the spinal cord requires WNT1 or WNT3A, our results are consistent with a model in which the relative concentration of WLS to Wnt regulates WNT1/3A signaling in the developing spinal cord.
Insights
Overexpressing WNTLESS (WLS) protein inhibits Wnt/β-catenin signaling in the developing spinal cord. The WLS to Wnt ratio appears to regulate WNT1/3A signaling during development.
Area of Science:
- Developmental biology
- Cell signaling
- Molecular biology
Background:
- WNTLESS (WLS) facilitates Wnt ligand transport from the Golgi to the cell surface.
- WLS loss-of-function studies in the CNS suggest roles in WNT1 and WNT3A signaling.
- Gain-of-function data for WLS has been limited.
Purpose of the Study:
- Investigate the phenotypic effects of WLS overexpression.
- Examine WLS function in cultured cells and the developing chick spinal cord.
- Clarify the role of WLS in Wnt/β-catenin pathway regulation.
Main Methods:
- Overexpression of WLS in HEK293T cells and chick spinal cord explants.
- Assessed Wnt/β-catenin pathway activity via cell proliferation and specification markers.
- Measured FZD10 expression as a Wnt-specific transcriptional readout.
Main Results:
- Low WLS levels with WNT1/WNT3A promoted Wnt/β-catenin signaling in HEK293T cells.
- High WLS levels inhibited Wnt/β-catenin signaling in both cell culture and spinal cord.
- WLS overexpression reduced FZD10 expression, indicating pathway inhibition.
Conclusions:
- WLS overexpression inhibits Wnt/β-catenin signaling in the developing spinal cord.
- Results support a model where WLS concentration relative to Wnt ligands regulates signaling.
- This regulation is critical for WNT1/3A signaling in spinal cord development.
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