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Brainstem

Submitted by: Nadia Villacampa, Universitat Autònoma de Barcelona 

The tissue in the image is a lesioned facial nucleus in the brainstem. A  peripheral injury was performed in the facial nerve of 3 month-old wild-type mice, the axon which leads to a retrograde neurodegeneration in the motorneurons of the facial nucleus was then resected. Among several changes in the facial motorneurons, they express IL10R (red) which leads to activation of the surrounding glial cells. In this case, reactive astrocytes are stained in green.

For the study of IL10R colocalization, sections were processed for IL10R staining as described in Villacampa N et al 2015, using Alexa Fluor® 555 conjugated streptavidin instead of horseradish streptavidin-peroxidase (1:1000; S-32355; Molecular Probes). The sections were then incubated overnight at 4°C and 1h at RT with mouse anti-GFAP (1:6000; 63893, Sigma-Aldrich) followed by Alexa Fluor® 488-conjugated anti-mouse secondary antibody(1:1000; A11029; Molecular Probes). Nuclei were counterstained with 4′,6-diamidino-2-phenylindole (DAPI; 1:10000; D9542; Sigma-Aldrich). Colocalization was analyzed with a Zeiss LSM 700 confocal microscope.

Magnification: 40X.  

 

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