Dermatan Sulfate-Free Mice Display Embryological Defects and Are Neonatal Lethal Despite Normal Lymphoid and Non-Lymphoid Organogenesis

Abstract

The epimerization of glucuronic acid into iduronic acid adds structural variability to chondroitin/dermatan sulfate polysaccharides. Iduronic acid-containing domains play essential roles in processes such as coagulation, chemokine and morphogen modulation, collagen maturation, and neurite sprouting. Therefore, we generated and characterized, for the first time, mice deficient in dermatan sulfate epimerase 1 and 2, two enzymes uniquely involved in dermatan sulfate biosynthesis. The resulting mice, termed DKO mice, were completely devoid of iduronic acid, and the resulting chondroitin sulfate chains were structurally different from the wild type chains, from which a different protein binding specificity can be expected. As a consequence, a vast majority of the DKO mice died perinatally, with greatly variable phenotypes at birth or late embryological stages such as umbilical hernia, exencephaly and a kinked tail. However, a minority of embryos were histologically unaffected, with apparently normal lung and bone/cartilage features. Interestingly, the binding of the chemokine CXCL13, an important modulator of lymphoid organogenesis, to mouse DKO embryonic fibroblasts was impaired. Nevertheless, the development of the secondary lymphoid organs, including the lymph nodes and spleen, was normal. Altogether, our results indicate an important role of dermatan sulfate in embryological development and perinatal survival.

Xyloside-primed Chondroitin Sulfate/Dermatan Sulfate from Breast Carcinoma Cells with a Defined Disaccharide Composition Has Cytotoxic Effects in Vitro

“…4 OAc, pH 8.0, containing 5 milliunits of each enzyme at 37°C for 16 h. Chondroitinase B degradations were performed in 25 l of 50 mM Tris-HCl, pH 7.5 (at 37°C), 50 mM NaCl, 4 mM CaCl 2 , 0.1 g/l BSA (Sigma) and 2 milliunits of enzyme at 37°C for 2 h. Heparinase I-III degradations were performed in 25 l of 20 mM HEPES, pH 7.2, 50 mM NaCl, 4 mM CaCl 2 , and 2.5 milliunits of each enzyme at 37°C for 16 h. CS A, CS B, CS C, and heparin were included as controls. Identification and quantification was performed by comparing the HPLC-separated 2-aminoacridone-labeled enzyme-generated disaccharides with 2-aminoacridone-labeled HPLC-separated disaccharide standards (Iduron) as previously described (22). The HS/CS/DS ratios of the XylNapOH-and XylNap-primed GAGs and the PG-GAGs were calculated as the relative proportions (% of total) of HS, IdoUA-containing disaccharides in CS/DS (CS/DS IdoUA ), and GlcUA-containing disaccharides in CS/DS (CS/DS GlcUA) according to Equations 2-4, respectively,…”

The Tyrosine Kinase Inhibitor Imatinib Augments Extracellular Fluid Exchange and Reduces Average Collagen Fibril Diameter in Experimental Carcinoma

“…GAG preparation, lyase treatment, fluorescence disaccharide labeling and separation were performed according to ref. 27. Briefly, tumors were protease and DNAase digested, and GAGs were purified on anion-exchange chromatography.…”

“…Fluorophore-labeling of the resulting disaccharides was performed by 2-aminoacridone (AMAC, Sigma). Pre-column AMAC-labeled disaccharides were analyzed with HPLC-fluorescence as described previously (27). Quantification was done by comparison to known weight of mock-treated standard disaccharides (Iduron).…”

Comparative Huntington and Parkinson Disease mRNA Analysis Reveals Common Inflammatory Processes

“…The most consistently ranked gene is RP1-93H18.7 (ENSG00000272403.1), a lncRNA, which was removed from Ensembl starting at version GRCh38.p2, but shows consistent transcription in these data. This gene is directly downstream of the gene DSE (dermatan sulfate epimerase) which is also DE in both HD and PD, is involved in embryonic development 20,21 , and has been related to the immune response in cancer patients 22 . Deficiencies in the second ranked gene, SPR (sepiapterin reductase), have been linked to DOPA-responsive dystonia 23 and previously implicated in PD 24 .…”