Spatial and temporal changes in the pattern of glycosylation of the developing chick limb tissue components as revealed by fluorescent conjugated lectin probes

Abstract

The changing pattern of expression of glycoconjugates during the differentiation of the chick leg bud between stages 17 to 34 (days 3 to 8 of incubation) was studied using fluorochrome-labelled plant lectins. Limb buds were fixed in cold acetic-alcohol and wax-embedded. Agglutinins of peanut (PNA), soybean (SBA) and succinylated wheat germ (WGAs) revealed a specific binding pattern in the apical ectodermal ridge (AER) between Hamburger and Hamilton stages 19–32. These stages coincide with the period of elevation of the AER. This specific binding pattern was absent from the adjacent dorsal and ventral ectoderm. Prechondrogenic cells were positive for WGA and for PNA, and the PNA-binding capacity was intensified after neuraminidase treatment. Premyogenic cells at stage 23 can be identified as negative to PNA after neuraminidase, while the blood vessels became positive. PNA, SBA, WGA, WGAs and, in addition, Ricinus communis (RCA-I) lectins stained the basal membrane. Strands of extracellular matrix which connect with the basal membrane and cross the limb transversely between dorsal and ventral ectoderm were stained by RCA-I, SBA and PNA after neuraminidase.