Human GLUT5 immunolabeling is useful for evaluating microglial status in neuropathological study using paraffin...

Area of Science:

• Neuropathology
• Neuroscience
• Immunohistochemistry

Background:

• Detecting resting and reactive microglia is crucial in neuropathological studies.
• Existing microglial markers have limitations in paraffin-embedded tissues.
• A need exists for a reliable microglial marker suitable for routine histopathology.

Purpose of the Study:

• To evaluate the utility of human glucose transporter 5 (hGLUT5) as a microglial marker.
• To assess hGLUT5 antibody performance in various tissue fixation and preparation methods.
• To compare hGLUT5 with established microglial markers for diagnostic applications.

Main Methods:

• Production of a specific antibody against the C-terminal sequence of hGLUT5.
• Staining of human brain tissue sections (cryostat and paraffin-embedded) using the hGLUT5 antibody.
• Double immunofluorescence analysis with glial fibrillary acidic protein (GFAP).
• Comparison with other microglial markers: KP-1, KiM1p, CR3.43, and RCA-1.

Main Results:

• The hGLUT5 antibody consistently stained microglia in both cryostat and paraffin sections.
• Both resting and reactive microglia were stained, with reactive microglia showing more intense labeling.
• hGLUT5 labeling revealed detailed microglial processes, outperforming Ki-M1P and KP-1.
• hGLUT5 stained both resting and reactive microglia, unlike CR3.43 (reactive only) and RCA-1 (microvessels).
• Staining remained effective even after prolonged formalin fixation, with microwave pretreatment enhancing immunoreactivity.

Conclusions:

• Human glucose transporter 5 (hGLUT5) is a valuable and effective marker for identifying both resting and reactive microglia.
• The hGLUT5 antibody is suitable for routine histopathological staining procedures due to its robustness and morphological clarity.
• hGLUT5 offers advantages over existing markers for detailed microglial morphology assessment in neuropathology.