Reduced amount of sulfated disaccharides isolated from HS of mutant embryos
| . | IV-A . | IV-S . | II-A . | II-S . | III-S . | I-S . |
|---|---|---|---|---|---|---|
| Wild type | 59.1±0.8 | 16.5±0.8 | 11.5±0.8 | 3.3±0.1 | 5.3±0.2 | 4.4±0.8 |
| Ndst1-/- | 75.7±0.7 | 6.5±0.2 | 11.2±0.9 | 2.9±0.14 | 1.2±0.07 | 2.6±0.14 |
| Ndst1+/- | 62.3 | 13.9 | 12.2 | 3.1 | 4.4 | 4.1 |
| . | IV-A . | IV-S . | II-A . | II-S . | III-S . | I-S . |
|---|---|---|---|---|---|---|
| Wild type | 59.1±0.8 | 16.5±0.8 | 11.5±0.8 | 3.3±0.1 | 5.3±0.2 | 4.4±0.8 |
| Ndst1-/- | 75.7±0.7 | 6.5±0.2 | 11.2±0.9 | 2.9±0.14 | 1.2±0.07 | 2.6±0.14 |
| Ndst1+/- | 62.3 | 13.9 | 12.2 | 3.1 | 4.4 | 4.1 |
HS was isolated from wild-type, Ndst1-/- and Ndst1+/- embryos, and samples were digested with heparin lyases. The resulting disaccharides were analyzed by anion exchange HPLC. HS from Ndst1-/- embryos was undersulfated, resulting in a decrease in N-sulfated and 2-O-sulfated disaccharides, especially III-S and I-S. HS derived from heterozygous embryos showed only mild effects on sulfation. Values indicate the mean % of total disaccharide. Three pooled mutant, wild-type and heterozygous embryos were investigated in one experiment. Results for the wild-type and mutant embryos are the mean±s.d. from two independent experiments. The abbreviations used indicate the following disaccharides: IV-A (ΔUA1,4GlcNAc), II-A(ΔUA1,4GlcNAc-6S), IV-S (ΔUA1,4GlcNS), II-S(ΔUA1,4GlcNS-6S), III-S (ΔUA2S1,4GlcNS) and I-S(ΔUA2S1,4GlcNS-6S). Students t-test revealed statistical significance for the differences observed (P<0.001, P<0.002, P<0.045, P<0.001 and P<0.05 for the change in IV-A, IV-S, II-S, III-S and I-S between wild-type and mutant HS, respectively).