Related Experiment Videos
Multiple roles for activated LEF/TCF transcription complexes during hair follicle development and differentiation
1Howard Hughes Medical Institute, Department of Molecular Genetics and Cell Biology and Committee on Developmental Biology, The University of Chicago, Chicago, IL 60637, USA.
Abstract:
LEF/TCF DNA-binding proteins act in concert with activated beta -catenin, the product of Wnt signaling, to transactivate downstream target genes. To probe the role of activated LEF/TCF transcription factor complexes in hair follicle morphogenesis and differentiation, we engineered mice harboring TOPGAL, a beta -galactosidase gene under the control of a LEF/TCF and beta -catenin inducible promoter. In mice, TOPGAL expression was directly stimulated by a stabilized form of beta -catenin, but was also dependent upon LEF1/TCF3 in skin. During embryogenesis, TOPGAL activation occurred transiently in a subset of LEF1-positive cells of pluripotent ectoderm and underlying mesenchyme. Downgrowth of initiated follicles proceeded in the absence of detectable TOPGAL expression, even though LEF1 was still expressed. While proliferative matrix cells expressed the highest levels of Lef1 mRNAs, LEF1 concentrated in the precursor cells to the hair shaft, where TOPGAL expression was co-induced with hair-specific keratin genes containing LEF/TCF-binding motifs. LEF1 and TOPGAL expression ceased during catagen and telogen, but reappeared at the start of the postnatal hair cycle, concomitant with precortex formation. In contrast to hair shaft precursor cells, postnatal outer root sheath expressed TCF3, but not TOPGAL. TCF3 was also expressed in the putative follicle stem cells, and while TOPGAL was generally silent in this compartment, it was stimulated at the start of the hair cycle in a fashion that appeared to be dependent upon stabilization of beta -catenin. Taken together, our findings demonstrate that LEF1/TCF3 is necessary but not sufficient for TOPGAL activation, revealing the existence of positive and negative regulators of these factors in the skin. Furthermore, our findings unveil the importance of activated LEF/TCF complexes at distinct times in hair development and cycling when changes in cell fate and differentiation commitments take place.
Insights
LEF/TCF transcription factors, regulated by Wnt signaling, are crucial for hair follicle development. Their activation is necessary but not sufficient for hair shaft formation, indicating complex regulation during hair cycling.
Area of Science:
- Developmental Biology
- Molecular Biology
- Genetics
Background:
- LEF/TCF DNA-binding proteins and beta-catenin mediate Wnt signaling to control gene expression.
- Understanding the role of LEF/TCF complexes in hair follicle morphogenesis and differentiation is essential.
Purpose of the Study:
- To investigate the function of activated LEF/TCF transcription factor complexes during hair follicle development and cycling.
- To characterize the temporal and spatial expression patterns of LEF/TCF activity in the mouse hair follicle.
Main Methods:
- Engineered TOPGAL reporter mice to visualize LEF/TCF activity.
- Analyzed TOPGAL expression in conjunction with LEF1 and TCF3 expression during embryonic and postnatal hair development.
- Correlated LEF/TCF activity with hair-specific keratin gene expression.
Main Results:
- TOPGAL expression, indicating LEF/TCF activity, was transiently activated during embryogenesis and reappeared during the postnatal hair cycle, particularly in hair shaft precursors.
- LEF1 expression was detected in proliferative matrix cells and hair shaft precursors, while TCF3 was found in the outer root sheath and putative stem cells.
- TOPGAL activation in hair shaft precursors coincided with hair-specific keratin gene expression, but follicle downgrowth occurred without detectable TOPGAL.
- LEF/TCF activity was stimulated at the start of the hair cycle in stem cells, dependent on beta-catenin stabilization.
Conclusions:
- Activated LEF/TCF complexes, particularly LEF1, play critical roles at specific stages of hair follicle morphogenesis and cycling.
- LEF1/TCF3 are necessary but not sufficient for TOPGAL activation, highlighting the involvement of other regulatory factors.
- The study reveals the dynamic regulation of Wnt/beta-catenin/LEF/TCF signaling in controlling hair cell fate and differentiation.