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Fluoxetine regulates neurogenesis in vitro through modulation of GSK-3β/β-catenin signaling.

Hui J, Zhang J, Kim H, Tong C, Ying Q, Li Z, Mao X, Shi G, Yan J, Zhang Z, Xi G - Int. J. Neuropsychopharmacol. (2014)

Bottom Line: The overexpression of a stabilized β-catenin protein (ΔN89 β-catenin) significantly increased NPC proliferation, while inhibition of β-catenin expression in NPCs led to a significant decrease in the proliferation and reduced the proliferative effects induced by fluoxetine.The effects of fluoxetine-induced up-regulation of both phosphorylation of Ser9 on GSK-3β and nuclear β-catenin were significantly prevented by the 5-hydroxytryptamine-1A (5-HT1A) receptor antagonist WAY-100635.The results demonstrate that fluoxetine may increase neurogenesis via the GSK-3β/β-catenin signaling pathway that links postsynaptic 5-HT1A receptor activation.

View Article: PubMed Central - PubMed

Affiliation: Department of Critical Care Medicine, Wuxi People's Hospital of Nanjing Medical University, Wuxi, China (Drs Hui and Yan); Department of Neurology, Wuxi People's Hospital of Nanjing Medical University, Wuxi, China (Drs J Zhang, Li, Mao, Shi, and Xi); Eli and Edythe Broad Center for Regenerative Medicine and Stem Cell Research at USC, Department of Cell and Neurobiology, University of Southern California, Los Angeles, CA (Drs Kim, Tong, and Ying); Department of Neurology, Affiliated ZhongDa Hospital of Southeast University, Nanjing, China (Dr Z Zhang).

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Inhibition of β-catenin expression decreased NPC proliferation and reduced the proliferative effects induced by fluoxetine. NPCs were transfected with β-catenin siRNA or a fluorescein-conjugated control siRNA. (A) β-catenin protein expression was detected by Western blotting and indicated a decrease expression (B). Values represent means ± SD (n = 5). *p < 0.01 versus control siRNA group. (C) 48h after transfection, some of the cells (as noted) were treated with 1 μM fluoxetine for an additional 48h before their cell proliferation was measured by BrdU labeling. Analyses of variance revealed main effects for β-catenin siRNA and fluoxetine treatment on NPC proliferation [F (1, 18) = 75.13, p < 0.0001 for β-catenin siRNA; F (1, 18) = 20.74, p < 0.0001 for fluoxetine]. Values represent means ± SD (n = 5). *p < 0.01 versus control siRNA group; #p < 0.001 versus fluoxetine + control siRNA group. BrdU, 2 d, 5’-bromo-2-deoxy-uridine; NPCs, neural precursor cells; SD, standard deviation; siRNA,.
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Figure 6: Inhibition of β-catenin expression decreased NPC proliferation and reduced the proliferative effects induced by fluoxetine. NPCs were transfected with β-catenin siRNA or a fluorescein-conjugated control siRNA. (A) β-catenin protein expression was detected by Western blotting and indicated a decrease expression (B). Values represent means ± SD (n = 5). *p < 0.01 versus control siRNA group. (C) 48h after transfection, some of the cells (as noted) were treated with 1 μM fluoxetine for an additional 48h before their cell proliferation was measured by BrdU labeling. Analyses of variance revealed main effects for β-catenin siRNA and fluoxetine treatment on NPC proliferation [F (1, 18) = 75.13, p < 0.0001 for β-catenin siRNA; F (1, 18) = 20.74, p < 0.0001 for fluoxetine]. Values represent means ± SD (n = 5). *p < 0.01 versus control siRNA group; #p < 0.001 versus fluoxetine + control siRNA group. BrdU, 2 d, 5’-bromo-2-deoxy-uridine; NPCs, neural precursor cells; SD, standard deviation; siRNA,.

Mentions: Finally, we performed a knockdown of β-catenin in NPCs using β-catenin siRNA. The efficiency of the transfection averaged 41.00±0.70% (n = 40 fields in four plates) when we counted the green fluorescent (GFP-positive) cells after 4 days. Protein expression was detected by Western blot (Figure 6A), which indicated a decreased expression from 0.98±0.05-fold to 0.59±0.06-fold (p < 0.01, n = 5, Figure 6B). ANOVA revealed main effects for β-catenin siRNA and fluoxetine treatment on NPC proliferation [F (1, 18) = 75.13, p < 0.0001 for β-catenin siRNA; F (1, 18) = 20.74, p < 0.0001 for fluoxetine]. Post hoc tests indicated that down-regulating β-catenin in NPCs significantly decreased (p < 0.01, n = 5) the proportion of BrdU-positive cells to 33.40±4.51% compared with the control siRNA (49.00±4.53%) and reversed the effects of fluoxetine on NPC proliferation (40.60±4.93% vs 60.40±4.28%, p < 0.001, n = 5, Figure 6C).


Fluoxetine regulates neurogenesis in vitro through modulation of GSK-3β/β-catenin signaling.

Hui J, Zhang J, Kim H, Tong C, Ying Q, Li Z, Mao X, Shi G, Yan J, Zhang Z, Xi G - Int. J. Neuropsychopharmacol. (2014)

Inhibition of β-catenin expression decreased NPC proliferation and reduced the proliferative effects induced by fluoxetine. NPCs were transfected with β-catenin siRNA or a fluorescein-conjugated control siRNA. (A) β-catenin protein expression was detected by Western blotting and indicated a decrease expression (B). Values represent means ± SD (n = 5). *p < 0.01 versus control siRNA group. (C) 48h after transfection, some of the cells (as noted) were treated with 1 μM fluoxetine for an additional 48h before their cell proliferation was measured by BrdU labeling. Analyses of variance revealed main effects for β-catenin siRNA and fluoxetine treatment on NPC proliferation [F (1, 18) = 75.13, p < 0.0001 for β-catenin siRNA; F (1, 18) = 20.74, p < 0.0001 for fluoxetine]. Values represent means ± SD (n = 5). *p < 0.01 versus control siRNA group; #p < 0.001 versus fluoxetine + control siRNA group. BrdU, 2 d, 5’-bromo-2-deoxy-uridine; NPCs, neural precursor cells; SD, standard deviation; siRNA,.
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Figure 6: Inhibition of β-catenin expression decreased NPC proliferation and reduced the proliferative effects induced by fluoxetine. NPCs were transfected with β-catenin siRNA or a fluorescein-conjugated control siRNA. (A) β-catenin protein expression was detected by Western blotting and indicated a decrease expression (B). Values represent means ± SD (n = 5). *p < 0.01 versus control siRNA group. (C) 48h after transfection, some of the cells (as noted) were treated with 1 μM fluoxetine for an additional 48h before their cell proliferation was measured by BrdU labeling. Analyses of variance revealed main effects for β-catenin siRNA and fluoxetine treatment on NPC proliferation [F (1, 18) = 75.13, p < 0.0001 for β-catenin siRNA; F (1, 18) = 20.74, p < 0.0001 for fluoxetine]. Values represent means ± SD (n = 5). *p < 0.01 versus control siRNA group; #p < 0.001 versus fluoxetine + control siRNA group. BrdU, 2 d, 5’-bromo-2-deoxy-uridine; NPCs, neural precursor cells; SD, standard deviation; siRNA,.
Mentions: Finally, we performed a knockdown of β-catenin in NPCs using β-catenin siRNA. The efficiency of the transfection averaged 41.00±0.70% (n = 40 fields in four plates) when we counted the green fluorescent (GFP-positive) cells after 4 days. Protein expression was detected by Western blot (Figure 6A), which indicated a decreased expression from 0.98±0.05-fold to 0.59±0.06-fold (p < 0.01, n = 5, Figure 6B). ANOVA revealed main effects for β-catenin siRNA and fluoxetine treatment on NPC proliferation [F (1, 18) = 75.13, p < 0.0001 for β-catenin siRNA; F (1, 18) = 20.74, p < 0.0001 for fluoxetine]. Post hoc tests indicated that down-regulating β-catenin in NPCs significantly decreased (p < 0.01, n = 5) the proportion of BrdU-positive cells to 33.40±4.51% compared with the control siRNA (49.00±4.53%) and reversed the effects of fluoxetine on NPC proliferation (40.60±4.93% vs 60.40±4.28%, p < 0.001, n = 5, Figure 6C).

Bottom Line: The overexpression of a stabilized β-catenin protein (ΔN89 β-catenin) significantly increased NPC proliferation, while inhibition of β-catenin expression in NPCs led to a significant decrease in the proliferation and reduced the proliferative effects induced by fluoxetine.The effects of fluoxetine-induced up-regulation of both phosphorylation of Ser9 on GSK-3β and nuclear β-catenin were significantly prevented by the 5-hydroxytryptamine-1A (5-HT1A) receptor antagonist WAY-100635.The results demonstrate that fluoxetine may increase neurogenesis via the GSK-3β/β-catenin signaling pathway that links postsynaptic 5-HT1A receptor activation.

View Article: PubMed Central - PubMed

Affiliation: Department of Critical Care Medicine, Wuxi People's Hospital of Nanjing Medical University, Wuxi, China (Drs Hui and Yan); Department of Neurology, Wuxi People's Hospital of Nanjing Medical University, Wuxi, China (Drs J Zhang, Li, Mao, Shi, and Xi); Eli and Edythe Broad Center for Regenerative Medicine and Stem Cell Research at USC, Department of Cell and Neurobiology, University of Southern California, Los Angeles, CA (Drs Kim, Tong, and Ying); Department of Neurology, Affiliated ZhongDa Hospital of Southeast University, Nanjing, China (Dr Z Zhang).

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Related in: MedlinePlus