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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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Fluoxetine increased the proliferation of NPCs. (A) Typical neurosphere morphology of rat embryonic neural precursor cells maintained in growth medium. (B–C) Immunostaining of Nestin (green) and DAPI (blue) in NPCs. Scale bars = 20 μm. (D–F) Immunostaining of sox2 (green) and DAPI (blue) in NPCs. (G) For cell proliferation, NPCs were incubated for 2 d in the presence of increasing concentrations (0–20 μM) of fluoxetin. Values represent means ± standard deviation (n = 5). BrdU-positive cells and nuclei (DAPI) were labeled with red and blue. Scale bars = 20 μm. (H) Quantification of data. ANOVA revealed a main effect of treatment [F (5, 24) = 9.67, p < 0.0005]. *p < 0.01 versus control (0 μM); #p < 0.005 versus control. BrdU, 2 d, 5’-bromo-2-deoxy-uridine; DAPI, 4,6-diamidino-2-phenylindole; NPCs, neural precursor cells.
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Figure 1: Fluoxetine increased the proliferation of NPCs. (A) Typical neurosphere morphology of rat embryonic neural precursor cells maintained in growth medium. (B–C) Immunostaining of Nestin (green) and DAPI (blue) in NPCs. Scale bars = 20 μm. (D–F) Immunostaining of sox2 (green) and DAPI (blue) in NPCs. (G) For cell proliferation, NPCs were incubated for 2 d in the presence of increasing concentrations (0–20 μM) of fluoxetin. Values represent means ± standard deviation (n = 5). BrdU-positive cells and nuclei (DAPI) were labeled with red and blue. Scale bars = 20 μm. (H) Quantification of data. ANOVA revealed a main effect of treatment [F (5, 24) = 9.67, p < 0.0005]. *p < 0.01 versus control (0 μM); #p < 0.005 versus control. BrdU, 2 d, 5’-bromo-2-deoxy-uridine; DAPI, 4,6-diamidino-2-phenylindole; NPCs, neural precursor cells.

Mentions: NPC cultures revealed aggregations and formation of typical neurospheres (Figure 1A), which was demonstrated by the immunocytochemical detection of nestin and sox2, two markers of undifferentiated NPCs (Figure 1B and 1E). We assessed the effects of increasing concentrations (0.001–20 μM) of fluoxetine on proliferation by BrdU labelling (Figure 1G). ANOVA revealed a main effect of treatment [F (5, 24) = 9.67, p < 0.0005]. These dose-response experiments, summarized in Figure 1H, show that after exposure to fluoxetine for 48h, cell proliferation was significantly increased at a concentration of 1 μM (p < 0.01, n = 5), whereas the highest concentration used (20 μM) actually decreased cell proliferation (p < 0.005, n = 5). Quantification of the data revealed that the percentage of BrdU-positive cells increased from 56.4±3.21% in untreated control cells to 70.40±4.39% in 1 μM fluoxetine-treated cells, but decreased to 46.80±3.42% in 20 μM fluoxetine-treated cells (Figure 1H).


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)

Fluoxetine increased the proliferation of NPCs. (A) Typical neurosphere morphology of rat embryonic neural precursor cells maintained in growth medium. (B–C) Immunostaining of Nestin (green) and DAPI (blue) in NPCs. Scale bars = 20 μm. (D–F) Immunostaining of sox2 (green) and DAPI (blue) in NPCs. (G) For cell proliferation, NPCs were incubated for 2 d in the presence of increasing concentrations (0–20 μM) of fluoxetin. Values represent means ± standard deviation (n = 5). BrdU-positive cells and nuclei (DAPI) were labeled with red and blue. Scale bars = 20 μm. (H) Quantification of data. ANOVA revealed a main effect of treatment [F (5, 24) = 9.67, p < 0.0005]. *p < 0.01 versus control (0 μM); #p < 0.005 versus control. BrdU, 2 d, 5’-bromo-2-deoxy-uridine; DAPI, 4,6-diamidino-2-phenylindole; NPCs, neural precursor cells.
© Copyright Policy - creative-commons
Related In: Results  -  Collection

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Figure 1: Fluoxetine increased the proliferation of NPCs. (A) Typical neurosphere morphology of rat embryonic neural precursor cells maintained in growth medium. (B–C) Immunostaining of Nestin (green) and DAPI (blue) in NPCs. Scale bars = 20 μm. (D–F) Immunostaining of sox2 (green) and DAPI (blue) in NPCs. (G) For cell proliferation, NPCs were incubated for 2 d in the presence of increasing concentrations (0–20 μM) of fluoxetin. Values represent means ± standard deviation (n = 5). BrdU-positive cells and nuclei (DAPI) were labeled with red and blue. Scale bars = 20 μm. (H) Quantification of data. ANOVA revealed a main effect of treatment [F (5, 24) = 9.67, p < 0.0005]. *p < 0.01 versus control (0 μM); #p < 0.005 versus control. BrdU, 2 d, 5’-bromo-2-deoxy-uridine; DAPI, 4,6-diamidino-2-phenylindole; NPCs, neural precursor cells.
Mentions: NPC cultures revealed aggregations and formation of typical neurospheres (Figure 1A), which was demonstrated by the immunocytochemical detection of nestin and sox2, two markers of undifferentiated NPCs (Figure 1B and 1E). We assessed the effects of increasing concentrations (0.001–20 μM) of fluoxetine on proliferation by BrdU labelling (Figure 1G). ANOVA revealed a main effect of treatment [F (5, 24) = 9.67, p < 0.0005]. These dose-response experiments, summarized in Figure 1H, show that after exposure to fluoxetine for 48h, cell proliferation was significantly increased at a concentration of 1 μM (p < 0.01, n = 5), whereas the highest concentration used (20 μM) actually decreased cell proliferation (p < 0.005, n = 5). Quantification of the data revealed that the percentage of BrdU-positive cells increased from 56.4±3.21% in untreated control cells to 70.40±4.39% in 1 μM fluoxetine-treated cells, but decreased to 46.80±3.42% in 20 μM fluoxetine-treated cells (Figure 1H).

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