The goal of the research was to investigate the microglial changes during retinal degeneration in P23H mice. Specifically, the study tries to show that the degeneration of photoreceptors affects the cells around those cells.
26 P23H rats were used in the study along with 26 control rats. This would enable the researchers to study the
Protein expression was analyzed using immunoblotting. The protein was run on a membrane then the membrane was probed using 43 different antibodies The detection were then performed. The immunoblotting analysis would indicate the amount and the level of different proteins that are available in the retina. The result of the immunoblotting are shown in figure 5, 7
The mice were euthanized and the eyes were enucleated and fixed in paraformaldehyde for one hour and cryoprotected in sucrose. The eyes were then dissected and the eyecups were processed and frozen. Then the sections were cut and mounted on a slide. This will allow for the image processing of the retina of both the P23H mice and the control mice, allowing for comparison. Then Retinal immunofluorescence was done on the mice, using the same antibodies and lectins. The previous sections were immunostained using the GFAP antibody, Gt antibody, Cx43 antibody, GS-IB4, and GAPDH antibody. These antibodies stain certain proteins in the retina and as a result, allow for a visualization of the slide that is obtained previously. The slides that were stained with the antibodies were analyzed for astrocytes the GFAPand TO-PRO-3 was used to identify astrocytes while the lectin was used to measure blood vessels astrocytes were counted manually, which allow for the examination of how much astrocytes there were in the retina. The result of histology are shown in figure 1,2,3,4,5,6,8,9.
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