Expression and functional involvement of N-cadherin in embryonic limb chondrogenesis

Abstract

Cell adhesion molecules have been shown to be important mediators of morphogenesis and pattern formation. In this study, we have shown that N-cadherin is expressed in a specific spatiotemporal manner in the developing limb bud during chondrogenesis in vivo and in cultured limb mesenchyme in vitro. The time period of maximal expression of N-cadherin corresponds to the period of active cellular condensation, an event believed to be a necessary prerequisite for chondrogenic differentiation. To directly assess the functional involvement of N-cadherin in cellular condensation, we have examined the effects of perturbing N-cadherin activity on both cell aggregation and chondrogenesis using NCD-2, a rat monoclonal antibody directed against the binding region of N-cadherin. Non-immune rat IgG was used as a control. Our results show that functional N-cadherin is necessary for chondrogenesis to proceed both in vivo and in vitro. Limb mesenchymal cells exhibited characteristic Ca(2+)-dependent cell aggregation in suspension, which was inhibited in the presence of exogenous NCD-2. In micromass cultures of limb mesenchymal cells, NCD-2 inhibited overt chondrogenesis in a dose-dependent manner. Furthermore, NCD-2 inhibition of chondrogenesis in micromass cultures was time-dependent, suggesting that N-cadherin is crucially involved during the latter half of the first 24 hours of culture, a time period most likely corresponding to active cellular condensation. NCD-2 also significantly influenced limb development when injected into embryonic limb buds in vivo. In addition to significant inhibition of chondrogenesis and developmental delays, gross developmental deformities and perturbation of overall pattern formation were also observed. Taken together, these results demonstrate that N-cadherin is functionally required in mediating the cell-cell interactions among mesenchymal cells important for chondrogenesis in micromass culture in vitro and in the intact limb bud in vivo.

Spatiotemporal Immunomodulation Using Biomimetic Scaffold Promotes Endochondral Ossification‐Mediated Bone Healing

“…The first factor taken into account is the function of transmembrane proteins, which is responsible for the interaction between cells and extracellular matrix. N cadherin is essential for initiating MSC condensation and negatively affects the Wnt/ β catenin signaling pathway, [ 32,33 ] while Integrin α 2 β 1 favors cartilage matrix condensation. [ 34 ] Therefore, the upregulation of both Integrin α 2 β 1 and N cadherin should contribute to early‐stage MSCs and cartilage matrix condensation under the unique immune microenvironment of B6D/M (Figure 4C).…”

Crosstalk of Endothelial and Mesenchymal Stromal Cells under Tissue-Related O2

“…Cadherin-cadherin contacts are known to provide a connection between the cytoskeleton components of neighboring cells [68][69][70][71]. A high level of N-cadherin expression is assumed to be characteristic of low committed stromal progenitors [72][73][74].…”

Potential Therapeutic Applications of N-Cadherin Antagonists and Agonists

“…These and other observations have given rise to the hypothesis that N-cadherin antagonists may be useful anti-cancer drugs (see next section). Furthermore, Mabs directed against N-cadherin block the differentiation of cells, such as osteoblasts (Hay et al, 2000;Marie 2002) and mesenchymal stem cells (MSC) (Oberlender and Tuan 1994;Tuan 2003) underlining the ability of CAMs to modulate gene expression. Short, synthetic linear peptides containing the Type I cadherin CAR sequence, His-Ala-Val (AHAVSE and LRAHAVDVNG) were then shown to be capable of disrupting N-cadherinmediated neurite outgrowth on astrocytes (Blaschuk et al, 1990b;Chuah et al, 1991).…”

“…These and other observations have given rise to the hypothesis that N-cadherin antagonists may be useful anti-cancer drugs (see next section). Furthermore, Mabs directed against N-cadherin block the differentiation of cells, such as osteoblasts ( Hay et al, 2000 ; Marie 2002 ) and mesenchymal stem cells (MSC) ( Oberlender and Tuan 1994 ; Tuan 2003 ) underlining the ability of CAMs to modulate gene expression.…”