Fabry disease is an X-linked lysosomal storage disease due to deficient α-galactosidase A (α-Gal A) activity and the resultant lysosomal accumulation of globotriaosylceramide (Gb3) and related lipids primarily in blood vessels, kidney, heart, and other organs. The renal distribution of stored glycolipid species in the α-Gal A knockout mouse model was compared to that in mice to assess relative distribution and absolute amounts of accumulated sphingolipid isoforms. Twenty isoforms of five sphingolipid groups were visualized by mass spectrometry imaging (MSI), and their distribution was compared with immunohistochemical (IHC) staining of Gb3, the major stored glycosphingolipid in consecutive tissue sections. Quantitative bulk lipid analysis of tissue sections was assessed by electrospray ionization with tandem mass spectrometry (ESI-MS/MS). In contrast to the findings in wild-type mice, all three analytical techniques (MSI, IHC, and ESI-MS/MS) revealed increases in Gb3 isoforms and ceramide dihexosides (composed mostly of galabiosylceramides), respectively. To our knowledge, this is the first report of the distribution of individual molecular species of Gb3 and galabiosylceramides in kidney sections in Fabry disease mouse. In addition, the spatial distribution of ceramides, ceramide monohexosides, and sphingomyelin forms in renal tissue is presented and discussed in the context of their biosynthesis.
- MeSH
- alfa-galaktosidasa genetika metabolismus MeSH
- Fabryho nemoc enzymologie genetika metabolismus MeSH
- hmotnostní spektrometrie MeSH
- imunochemie MeSH
- ledviny chemie metabolismus MeSH
- lidé MeSH
- modely nemocí na zvířatech MeSH
- myši knockoutované MeSH
- myši MeSH
- sfingolipidy chemie metabolismus MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Většina pokusů studujících strukturu či dynamiku biomakromolekul je prováděna in vitro z důvodu vysokých požadavků na čistotu a koncentraci vzorku. Metoda in-cell NMR ve spojení se izotopovým značením a molekulární genetikou nabízí nástroj, jak studovat biomakromolekuly v jejich přirozeném prostředí - v buňce. Jsme tak schopni získat informace o posttranslačních modifikacích, o fysiologických či patologických konfirmacích proteinu na atomární úrovni, nebo o dynamice vazby ligandů. Otevírá se tak značné množství nových badatelských oblastí pro molekulární biologii, biochemii či medicínu a farmakologii.
Most experiments studying the structure and dynamics of biomacromolecules are performed in vitro because of high requirements for purity and concetration. In-cell NMR in connection to isotope labeling and molecular genetics offers a great tool for study of biomacromolecules in their natural environment - in the cell. Thus, we are able to obtain informations about post-translational modrtications, about physiological or patological conformations of protein, or dynamic binding of the ligands, all at the atomic level. Large number of new area is opened for molecular biology, biochemistry, medicine and farmacology research.
