Expression of multiple CD44 isoforms in the apical ectodermal ridge of the embryonic mouse limb

Previous immunohistochemical studies have shown that CD44 is highly enriched within the apical ectodermal ridge of the developing limb (Wheatley et al. [1993] Development 119: 295-306), but the particular isoforms of CD44 were not identified. We show here that CD44s (standard or "hemopoietic" isoform) and several CD44 variants, especially V3-V10, V4-V10, and V6-V10, are concentrated in the apical ectodermal ridge in the early mouse limb. Since CD44s is a major cell surface receptor for hyaluronan, we compared its localization with that of hyaluronan. In the early limb bud, hyaluronan is distributed throughout the mesoderm but is absent from all regions of the ectoderm. Hyaluronan is especially enriched in the basement membrane separating ectoderm and mesoderm, except beneath the apical ectodermal ridge where it is absent. Since CD44s is a known endocytic receptor for hyaluronan, its presence in ridge ectoderm could lead to degradation of hyaluronan destined for the neighboring region of basement membrane, thus facilitating interaction of the ridge with underlying mesoderm. The CD44 (V3-V10) isoform found in the ridge is expressed elsewhere as a proteoglycan with heparan sulfate chains that bind fibroblast growth factors. Since fibroblast growth factors are present in the ridge and are essential for limb morphogenesis, CD44 (V3-V10) is likely to act as a cofactor or modulator in the growth-promoting action or maintenance of the ridge.