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Fig. 8 | Molecular Neurodegeneration

Fig. 8

From: Functional microRNA targetome undergoes degeneration-induced shift in the retina

Fig. 8

AGO2 gene and protein expression changes following PD. (A) Diagram indicating retinal regions where histological measures were taken. Lesion site refers to the area most damaged by PD. (B) (i) AGO2 in situ hybridization in dim and PD retinas at the three retinal sites indicated in A. AGO2 mRNA increased at all three locations and is distributed across the GCL, INL in the inferior retina and lesion site. In the superior retina, AGO2 mRNA expression is also visible within the ONL (yellow arrow) and the OLM (orange arrow). (ii) Co-labelling of AGO2 mRNA and glutamine synthetase in the superior retina showing colocalization along the OLM and Müller glia processes. Gray micrographs are DIC images of AGO2 in situ hybridization (as shown in i). Red labelling are pseudo-coloured pixels corresponding to the area positive for AGO2 labelling (dark spots in the adjacent DIC image). (C) (i) Image analysis pipeline used for colocalization analysis. (ii) Immunohistochemistry showed AGO2 protein expression is confined within the GCL, INL and photoreceptor segments in dim retinas. Following PD, AGO2 protein expression is elevated within the INL and photoreceptor segments. Co-localization of AGO2 labelling with Müller glia visualized using a reporter mouse strain showed distinct colocalization along the inferior and superior OLM (inset 2), Müller glia processes (inset 4) and Müller glia cell bodies (inset 3). In the absence of PD limited colocalization is seen in the superior retina (inset 1). (D) (i) Axis description for plots shown in ii. (ii) AGO2 protein expression profile quantified across retinal layers and depicted as pixel intensity (y-axis), along the outer to inner retinal direction (green x-axis) and superior to inferior (orange z-axis). (E) (i) AGO2 western blot and corresponding GAPDH loading control. (ii) GAPDH-normalized densitometry values for AGO2 (Error bars show SEM, * P < 0.05, Student’s t-test)

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