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Updated: Apr 23, 2026
Chicken Recombinant Limbs Assay to Understand Morphogenesis, Patterning, and Early Steps in Cell Differentiation
Published on: January 12, 2022
Effect of FGFR inhibitors on chicken limb development
Dana Horakova1, Petra Cela, Pavel Krejci
1Department of Anatomy, Histology and Embryology, Faculty of Veterinary Medicine, University of Veterinary and Pharmaceutical Sciences, Brno, Czech Republic.
Abstract:
Fibroblast growth factor (FGF) signalling appears essential for the regulation of limb development, but a full complexity of this regulation remains unclear. Here, we addressed the effect of three different chemical inhibitors of FGF receptor tyrosine kinases (FGFR) on growth and patterning of the chicken wings. The inhibitor PD173074 caused shorter and thinner wing when using lower concentration. Microinjection of higher PD173074 concentrations (25 and 50 mmol/L) into the wing bud at stage 20 resulted in the development of small wing rudiment or the total absence of the wing. Skeletal analysis revealed the absence of the radius but not ulna, deformation of metacarpal bones and/or a reduction of digits. Treatment with PD161570 resembled the effects of PD173074. NF449 induced shortening and deformation of the developing wing with reduced autopodium. These malformed embryos mostly died at the stage HH25-29. PD173074 reduced chondrogenesis also in the limb micromass cultures together with early inhibition of cartilaginous nodule formation, evidenced by lack of sulphated proteoglycan and peanut agglutinin expression. The effect of FGFR inhibition on limb development observed here was unlikely mediated by excessive cell death as none of the inhibitors caused massive apoptosis at low concentrations. More probably, FGFR inhibition decreased both the proliferation and adhesion of mesenchymal chondroprogenitors. We conclude that FGFR signalling contributes to the regulation of the anterior-posterior patterning of zeugopod during chicken limb development.
Insights
Fibroblast growth factor receptor (FGFR) inhibition disrupts chicken wing development, causing malformations and reduced growth. This suggests FGFR signaling is crucial for limb patterning and chondrogenesis.
Area of Science:
- Developmental Biology
- Molecular Biology
- Cell Biology
Background:
- Fibroblast growth factor (FGF) signaling is vital for limb development.
- The precise regulatory mechanisms of FGF signaling in limb development are not fully understood.
Purpose of the Study:
- To investigate the effects of three chemical inhibitors of FGF receptor tyrosine kinases (FGFR) on chicken wing growth and patterning.
- To elucidate the role of FGFR signaling in limb development and chondrogenesis.
Main Methods:
- Chemical inhibition of FGFR using PD173074, PD161570, and NF449 in chicken embryos.
- Microinjection of inhibitors into wing buds at specific developmental stages.
- Skeletal analysis of limb malformations.
- Limb micromass cultures to assess chondrogenesis.
- Apoptosis assays to evaluate cell death.
Main Results:
- PD173074 and PD161570 caused dose-dependent wing shortening, thinning, and malformations, including digit reduction and absence of the radius.
- Higher concentrations of PD173074 led to severe wing defects or complete absence.
- NF449 induced wing shortening, deformation, and autopodium reduction, with high embryonic mortality.
- FGFR inhibition reduced chondrogenesis and cartilaginous nodule formation in micromass cultures.
- Inhibitors did not cause significant apoptosis, suggesting reduced proliferation and adhesion of chondroprogenitors.
Conclusions:
- FGFR signaling is essential for normal chicken wing growth, patterning, and chondrogenesis.
- Inhibition of FGFR signaling leads to limb malformations by affecting mesenchymal chondroprogenitor proliferation and adhesion.
- FGFR signaling regulates anterior-posterior patterning of the zeugopod during chicken limb development.
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