GNE deficiency impairs Myogenesis in C2C12 cells and cannot be rescued by ManNAc supplementation.

GNE myopathy (GNEM) is a late-onset muscle atrophy, caused by mutations in the gene for the key enzyme of sialic acid biosynthesis, UDP-N-acetylglucosamine 2-epimerase/N-acetylmannosamine kinase (GNE). With an incidence of one to nine cases per million it is an ultra-rare, so far untreatable, autosomal recessive disease. Several attempts have been made to treat GNEM patients by oral supplementation with sialic acid precursors (e.

Evaluation of -Acetylmannosamine Administration to Restore Sialylation in -Deficient Human Embryonal Kidney Cells.

Background: A key mechanism in the neuromuscular disease GNE myopathy (GNEM) is believed to be that point mutations in the gene impair sialic acid synthesis - maybe due to UDP--acetylglucosamine 2-epimerase/-acetylmannosamine kinase (GNE) activity restrictions - and resulting in muscle tissue loss. -acetylmannosamine (ManNAc) is the first product of the bifunctional GNE enzyme and can therefore be regarded as a precursor of sialic acids. This study investigates whether this is also a suitable substance for restoring the sialic acid content in -deficient cells.

Glycation Interferes with the Activity of the Bi-Functional UDP--Acetylglucosamine 2-Epimerase/-Acetyl-mannosamine Kinase (GNE).

Mutations in the gene coding for the bi-functional UDP--acetylglucosamine 2-epimerase/-acetylmannosamine kinase (GNE), the key enzyme of the sialic acid biosynthesis, are responsible for autosomal-recessive GNE myopathy (GNEM). GNEM is an adult-onset disease with a yet unknown exact pathophysiology. Since the protein appears to work adequately for a certain period of time even though the mutation is already present, other effects appear to influence the onset and progression of the disease.

Variations in Plasma Membrane Topography Can Explain Heterogenous Diffusion Coefficients Obtained by Fluorescence Correlation Spectroscopy.

Fluorescence correlation spectroscopy (FCS) is frequently used to study diffusion in cell membranes, primarily the plasma membrane. The diffusion coefficients reported in the plasma membrane of the same cell type and even within single cells typically display a large spread. We have investigated whether this spread can be explained by variations in membrane topography throughout the cell surface, that changes the amount of membrane in the FCS focal volume at different locations.

Correlative Stimulated Emission Depletion and Scanning Ion Conductance Microscopy.

Correlation microscopy combining fluorescence and scanning probe or electron microscopy is limited to fixed samples due to the sample preparation and nonphysiological imaging conditions required by most probe or electron microscopy techniques. Among the few scanning probe techniques that allow imaging of living cells under physiological conditions, scanning ion conductance microscopy (SICM) has been shown to be the technique that minimizes the impact on the investigated sample. However, combinations of SICM and fluorescence microscopy suffered from the mismatch in resolution due to the limited resolution of conventional light microscopy.

A low-cost, large field-of-view scanning ion conductance microscope for studying nanoparticle-cell membrane interactions.

Nanoparticles have the potential to become versatile tools in the medical and life sciences. One potential application is delivering drugs or other compounds to the cell cytoplasm, which requires the nanoparticles to bind to or cross the cell membrane. However, there are only a few tools available which allow studying the interaction of nanoparticles and the cell membrane of living cells in a physiological environment.

Long-term, long-distance recording of a living migrating neuron by scanning ion conductance microscopy.

Bias-free, three-dimensional imaging of entire living cellular specimen is required for investigating shape and volume changes that occur during cellular growth or migration. Here we present fifty consecutive recordings of a living cultured neuron from a mouse dorsal root ganglion obtained by Scanning ion conductance microscopy (SICM). We observed a saltatory migration of the neuron with a mean velocity of approximately 20 μm/h.