Long‐term in vitro analysis of limb cartilage development: Involvement of Wnt signaling
- 26 July 2004
- journal article
- research article
- Published by Wiley in Journal of Cellular Biochemistry
- Vol. 93 (3), 526-541
- https://doi.org/10.1002/jcb.20190
Abstract
Endochondral skeletal development involves the condensation of mesenchymal cells, their differentiation into chondrocytes, followed by chondrocyte maturation, hypertrophy, and matrix mineralization, and replacement by osteoblasts. The Wnt family of secreted proteins have been shown to play important roles in vertebrate limb formation. To examine the role(s) of Wnt members and their transmembrane-spanning receptor(s), Frizzled (fz), we retrovirally misexpressed Wnt-5a, Wnt-7a, chicken frizzled-1 (Chfz-1), and frizzled-7 (Chfz-7) in long-term (21 day) high density, micromass cultures of stage 23/24 chick embryonic limb mesenchyme. This culture system recapitulates in vitro the entire differentiation (days 1–10), growth (days 5–12), and maturation and hypertrophy (from day 12 on) program of cartilage development. Wnt-7a misexpression severely inhibited chondrogenesis from day 7 onward. Wnt-5a misexpression resulted in a poor hypertrophic phenotype by day 14. Chfz-7 misexpression caused a slight delay of chondrocyte maturation based on histology, whereas Chfz-1 misexpression did not affect the chondrogenic phenotype. Misexpression of all Wnt members decreased collagen type X expression and alkaline phosphatase activity at day 21. Our findings implicate functional role(s) for Wnt signaling throughout embryonic cartilage development, and show the utility of the long-term in vitro limb mesenchyme culture system for such studies.Keywords
This publication has 48 references indexed in Scilit:
- Wnt regulation of limb mesenchymal chondrogenesis is accompanied by altered N‐cadherin‐related functionsThe FASEB Journal, 2001
- Mechanism and function of signal transduction by the Wnt/β-catenin and Wnt/Ca2+ pathwaysOncogene, 1999
- MECHANISMS OF WNT SIGNALING IN DEVELOPMENTAnnual Review of Cell and Developmental Biology, 1998
- Wnt Signaling Stabilizes the Dual‐Function Protein β‐Catenin in Diverse Cell TypesaAnnals of the New York Academy of Sciences, 1998
- Modulation of Embryonic Intracellular Ca2+Signaling byWnt-5ADevelopmental Biology, 1997
- A series of normal stages in the development of the chick embryoDevelopmental Dynamics, 1992
- Identification of programmed cell death in situ via specific labeling of nuclear DNA fragmentation.The Journal of cell biology, 1992
- The role of Wnt genes in vertebrate developmentCurrent Opinion in Genetics & Development, 1992
- Chondrogenesis of limb bud mesenchyme in vitro: Stimulation by cationsDevelopmental Biology, 1986
- Stage-related capacity for limb chondrogenesis in cell cultureDevelopmental Biology, 1977