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Sensitive quantification of α-glucans in mouse tissues, cell cultures, and human cerebrospinal fluid
- Source :
- J Biol Chem
- Publication Year :
- 2020
-
Abstract
- The soluble α-polyglucan glycogen is a central metabolite enabling transient glucose storage to suit cellular energy needs. Glycogen storage diseases (GSDs) comprise over 15 entities caused by generalized or tissue-specific defects in enzymes of glycogen metabolism. In several, e.g. in Lafora disease caused by the absence of the glycogen phosphatase laforin or its interacting partner malin, degradation-resistant abnormally structured insoluble glycogen accumulates. Sensitive quantification methods for soluble and insoluble glycogen are critical to research, including therapeutic studies, in such diseases. This paper establishes methodological advancements relevant to glycogen metabolism investigations generally, and GSDs. Introducing a pre-extraction incubation method, we measure degradation-resistant glycogen in as little as 30 mg of skeletal muscle or a single hippocampus from Lafora disease mouse models. The digestion-resistant glycogen correlates with the disease-pathogenic insoluble glycogen and can readily be detected in very young mice where glycogen accumulation has just begun. Second, we establish a high-sensitivity glucose assay with detection of ATP depletion, enabling 1) quantification of α-glucans in cell culture using a medium-throughput assay suitable for assessment of candidate glycogen synthesis inhibitors, and 2) discovery of α-glucan material in healthy human cerebrospinal fluid, establishing a novel methodological platform for biomarker analyses in Lafora disease and other GSDs.
- Subjects :
- 0301 basic medicine
Male
Metabolite
Phosphatase
Cell Culture Techniques
Biochemistry
Hippocampus
Lafora disease
03 medical and health sciences
chemistry.chemical_compound
Mice
medicine
Glycogen storage disease
Animals
Humans
Glycogen synthase
Muscle, Skeletal
Molecular Biology
Glucans
030102 biochemistry & molecular biology
Glycogen
biology
Chemistry
Molecular Bases of Disease
Cell Biology
medicine.disease
Glycogen Storage Disease
Mice, Inbred C57BL
030104 developmental biology
HEK293 Cells
Lafora Disease
Cell culture
biology.protein
Female
Laforin
Subjects
Details
- ISSN :
- 1083351X
- Volume :
- 295
- Issue :
- 43
- Database :
- OpenAIRE
- Journal :
- The Journal of biological chemistry
- Accession number :
- edsair.doi.dedup.....f5d53d94f0a3823eb1fe585caf8bc01a