engleski

Gangliosides in cerebrospinal fluid: tackling the optimal method of extraction, purification and characterization of gangliosides as potential diagnostic biomarkers in neurological disorders

Gangliosides are membrane lipids, found with greatest abundance and structural variability in human brain tissue. Importance of gangliosides in various cellular and physiological processes has been well established, with particular emphasis on their roles in establishing proper axon-glial interactions, maintenance of axon stability, axon regeneration, and modulation of nerve cell excitability. In addition, there is strong evidence on the involvement of gangliosides in neurodegeneration pathogenesis. Despite continuous effort in investigation of neurodegenerative biomarkers in clinically available samples, to date a disappointingly small number of specific diagnostic markers in cerebrospinal fluid (CSF) have been determined. This fact explains the interest in compositional analysis of gangliosides in cerebrospinal fluid as potential biomarkers for various neurological disorders. However, there are several major limitations for analysis of CSF gangliosides. In comparison with brain tissue, ganglioside concentration in CSF is extremely low. Furthermore, association of gangliosides and proteins in CSF due to formation of protein-ganglioside divalent cation complexes has been described. These intermolecular interactions make successful CSF ganglioside analysis much more demanding, especially since pathological CSF samples contain larger amounts of proteins. The aim of our study was to establish an optimal protocol for extraction, purification and characterization of CSF gangliosides with long- term goal of utilizing that protocol in research and diagnostics of different neurological disorders. Preliminary analysis was performed in CSF samples derived from patients with clinical diagnosis of Alzheimer’s disease and mild-cognitive impairment in comparison with control CSF samples. We employed different approaches for extraction, purification and chromatographic analysis of gangliosides in native CSF samples. We also implemented methodological modifications which enabled separation and consequent removal of protein-cation complexes from gangliosides. Finally, we used mass spectrometry for detailed structural and compositional analysis of CSF gangliosides. This combination of classical biochemical methods and sophisticated structural analysis provides a solid base for future investigation of CSF gangliosides. We believe that will prove to be a valuable tool in setting gangliosides as new CSF biomarkers in diverse neurological disorders.

Gangliosides; Cerebrospinal fluid; Neurodegeneration; Biomarkers

nije evidentirano