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Fig. 5 | Journal of Neuroinflammation

Fig. 5

From: Neurodegeneration in the centrally-projecting Edinger–Westphal nucleus contributes to the non-motor symptoms of Parkinson’s disease in the rat

Fig. 5

Selective partial UCN1 neuron ablation in the centrally-projecting Edinger-Wespthal nucleus (EWcp) causes increased anhedonia and anxiety level without affecting motor skills. Leptin–saporin-injected rats (red bars) show reduced preference to the sweetened water in the sucrose preference test (A). Open field test (OFT) revealed longer period of time spent in the peripheral part of the arena (B). Neither the total traveled distance in OFT (C) nor the rotarod test (D) revealed compromised motor performance of leptin-saporin-treated rats in comparison to saporin-injected controls (black bars). UCN1 (red)—NeuN (green) double labeling (E, F) revealed reduced UCN1/NeuN cell count upon leptin-saporin treatment (F, G), while the number of UCN1 immunonegative NeuN neurons remained unchanged in the EWcp (H). Ionized calcium binding adaptor molecule 1 (IBA1)-immunoreactive (ir) microglia (green in I and J) showed increased activity upon leptin-saporin treatment (J, K) and tended to interact with UCN1-ir cells more frequently (see the blue arrowheads in J and graph P). Astrocytes showing glial fibrillary acidic protein (GFAP, red in I and J) were significantly (L) more active upon leptin-saporin injection (J), although some active cells were also seen in saporin-treated controls (I) representing reactive gliosis due to the effect of surgical manipulation. UCN1-ir (green in M and N) neurons were seen to contain the apoptotic marker caspase 3 (see the boxed area and blue arrowhead in N) in leptin-saporin injected rats (O), while in saporin-injected control animals, only some glial nuclei were positive for caspase 3 (M). Saporin (Q) and leptin–saporin injection (R) into the EWcp did not affect the number of tyrosine-hydroxylase (TH, green in Q and R)-ir cells in the substantia nigra pars compacta (SNpc) as revealed by unbiased stereological cell counting (T). Caspase 3 (red in Q and R) revealed no remarkable apoptotic activity of SNpc/TH (S). Both microglia (IBA, green in U and V) and astroglial (GFAP) activity scores remained basal (W and X, respectively) in both groups (n = 6–9) t: tendency (p = 0.051), *p < 0.05, **p < 0.01, ***p < 0.001 in Student’s t test or in Mann–Whitney U test in K, L, W, X. ns not significant. Bars: 50 µm

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