Fig. 7From: Sustained hyperammonemia induces TNF-a IN Purkinje neurons by activating the TNFR1-NF-κB pathwayTNF-a is increased in glial cells but not in Purkinje neurons after 2 weeks of hyperammonemia. An increase in TNF-a was observed in glial cells (white matter, b, e) but not in Purkinje neurons (a, d). Double immunofluorescence confirmed the presence of TNF-a in microglia (c) and astrocytes (d), and the proportion of these cells expressing TNF-a was quantified (f). To investigate the contribution of the nuclear translocation of NF-kB to TNF-a synthesis, immunofluorescence for the p50 subunit was performed in the Purkinje layer, and representative images are shown in g (green staining) and quantified in j. Double immunofluorescence was also performed for NF-kB p50 (green in h and red in i) and Iba-1, a microglial marker (red in h) or GFAP, an astroglial marker (green in i). The merged image shows the colocalization of these proteins (yellow). The number of microglia and astrocytes expressing nuclear p50 are quantified in k and l, respectively. The values are the mean ± SEM of 3 rats per group. Values that were significantly different from those of the control rats are indicated by asterisks. *p ≤ 0.05, **p ≤ 0.051, ****p ≤ 0.0001. ++ refers to microglia expressing TNF-a in HA VH rats vs microglia expressing TNF-a in C VH rats (p < 0.01) and ## refers to astrocytes expressing TNF-a in HA VH rats vs astrocytes expressing TNF-a in C VH rats (p < 0.01). C VH = vehicle-treated control rats, HA VH = vehicle-treated hyperammonemic ratsBack to article page