Heparan sulfate (HS) binds numerous extracellular ligands, including cell-cell signaling molecules and their signal-transducing receptors. Ligand binding sites in HS have specific sulfation patterns; and several observations suggest that the HS sulfation pattern is the same for every HS chain that a cell synthesizes, regardless of the core protein to which it is attached. Nonetheless, virtually every Drosophila, zebrafish, Xenopus, and mouse that lacks a specific HS core protein has a mutant phenotype, even though other HS core proteins are expressed in the affected cells. Genetic manipulation of HS core protein genes is beginning to indicate that HS core proteins have functional specificities that are required during distinct stages of development.