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Neuron-Macrophage Co-cultures to Activate Macrophages Secreting Molecular Factors with Neurite Outgrowth Activity
Published on: March 30, 2018
Macrophage migration inhibitory factor promotes proliferation and neuronal differentiation of neural stem/precursor
Xijing Zhang1, Liangwei Chen, Yazhou Wang
11. Department of Anesthesiology, Xijing Hospital, the Fourth Military Medical University, Xi'an 710032, China.
Abstract:
Macrophage migration inhibitory factor (MIF) is a highly conserved and evolutionarily ancient mediator with pleiotropic effects. Recent studies demonstrated that the receptors of MIF, including CD44, CXCR2, CXCR4 and CD74, are expressed in the neural stem/progenitor cells (NSPCs). The potential regulatory effect of MIF on NSPCs proliferation and neuronal differentiation, however, is largely unknown. Here, we investigated the effect of MIF on NSPC proliferation and neuronal differentiation, and further examined the signal pathway by which MIF transduced these signal effects in mouse NSPCs in vitro. The results showed that both Ki67-positive cells and neurosphere volumes were increased in a dose-dependent manner following MIF treatment. Furthermore, the expression of nuclear β-catenin was significantly stronger in MIF-stimulated groups than that in control groups. Conversely, administration of IWR-1, the inhibitor of Wnt/β-catenin pathway, significantly inhibited the proliferative effect of MIF on NSPCs. Immunostaining and Western blot further indicated that doublecortin (DCX) and Tuj 1, two neuronal markers, were evidently increased with MIF stimulation during NSPC differentiation, and there were more Tuj1-positive cells migrated out from neurospheres in MIF-stimulated groups than those in control groups. During NSPC differentiation, MIF increased the activity of β-galactosidase that responds to Wnt/β-catenin signaling. Wnt1 and β-catenin proteins were also up-regulated with MIF stimulation. Moreover, the expression of DCX and Tuj 1 was inhibited significantly by IWR-1. Taken together, the present study indicated that MIF enhances NSPC proliferation and promotes the neuronal differentiation, by activating Wnt/β-catenin signal pathway. The interaction between MIF and Wnt/β-catenin signal pathway may play an important role in modulating NSPC renewal and fate during brain development.
Insights
Macrophage migration inhibitory factor (MIF) boosts neural stem cell proliferation and neuronal differentiation by activating the Wnt/β-catenin pathway. This finding highlights MIF
Area of Science:
- Neuroscience
- Cell Biology
- Developmental Biology
Background:
- Macrophage migration inhibitory factor (MIF) is a pleiotropic mediator with known roles in inflammation and immunity.
- MIF receptors (CD44, CXCR2, CXCR4, CD74) are expressed in neural stem/progenitor cells (NSPCs).
- The specific role of MIF in regulating NSPC behavior remains largely uncharacterized.
Purpose of the Study:
- To investigate the effect of MIF on the proliferation and neuronal differentiation of mouse NSPCs in vitro.
- To elucidate the underlying signaling pathway mediating MIF's effects on NSPCs.
Main Methods:
- Treatment of mouse NSPCs with varying concentrations of MIF.
- Assessment of NSPC proliferation using Ki67 staining and neurosphere formation assays.
- Analysis of neuronal differentiation markers (Doublecortin, Tuj 1) via immunostaining and Western blot.
- Pharmacological inhibition of the Wnt/β-catenin pathway using IWR-1.
- Measurement of Wnt/β-catenin signaling activity.
Main Results:
- MIF treatment dose-dependently increased NSPC proliferation, evidenced by elevated Ki67-positive cells and larger neurospheres.
- MIF stimulation significantly enhanced neuronal differentiation, indicated by increased Doublecortin (DCX) and Tuj 1 expression and migration.
- MIF activated the Wnt/β-catenin pathway, shown by increased nuclear β-catenin, Wnt1, and β-galactosidase activity.
- Inhibition of Wnt/β-catenin signaling with IWR-1 blocked MIF's proliferative and differentiation-promoting effects.
Conclusions:
- MIF promotes NSPC proliferation and enhances neuronal differentiation.
- These effects are mediated through the activation of the Wnt/β-catenin signaling pathway.
- The MIF-Wnt/β-catenin axis represents a crucial regulatory mechanism in neural stem cell renewal and fate determination during brain development.
