| Autor: |
Gramegna Tota C; Unit of Biochemistry, Department of Molecular Medicine, University of Pavia, 27100 Pavia, Italy., Leone A; Unit of Biochemistry, Department of Molecular Medicine, University of Pavia, 27100 Pavia, Italy.; University School for Advanced Studies Pavia, IUSS Pavia, 27100 Pavia, Italy., Khan A; Unit of Biochemistry, Department of Molecular Medicine, University of Pavia, 27100 Pavia, Italy., Forlino A; Unit of Biochemistry, Department of Molecular Medicine, University of Pavia, 27100 Pavia, Italy., Rossi A; Unit of Biochemistry, Department of Molecular Medicine, University of Pavia, 27100 Pavia, Italy., Paganini C; Centre for Inherited Diseases, Department of Research, Fondazione IRCCS Policlinico San Matteo, 27100 Pavia, Italy. |
| Abstrakt: |
Desbuquois dysplasia type 1 (DBQD1) is a recessive chondrodysplasia caused by mutations in the CANT1 gene, encoding for the Golgi Calcium-Activated Nucleotidase 1 (CANT1). The enzyme hydrolyzes UDP, the by-product of glycosyltransferase reactions, but it might play other roles in different cell types. Using a Cant1 knock-out mouse, we demonstrated that CANT1 is crucial for glycosaminoglycan (GAG) synthesis; however, its impact on the biochemical properties of cartilage proteoglycans remains unknown. Thus, in this work, we characterized decorin and aggrecan from primary chondrocyte cultures and cartilage biopsies of mutant mice at post-natal day 4 by Western blots and further investigated their distribution in the cartilage extracellular matrix (ECM) by immunohistochemistry. We demonstrated that the GAG synthesis defect caused by CANT1 impairment led to the synthesis and secretion of proteoglycans with shorter GAG chains compared with wild-type animals. However, this alteration did not result in the synthesis and secretion of decorin and aggrecan in the unglycanated form. Interestingly, the defect was not cartilage-specific since also skin decorin showed a reduced hydrodynamic size. Finally, immunohistochemical studies in epiphyseal sections of mutant mice demonstrated that the proteoglycan structural defect moderately affected decorin distribution in the ECM. |
