Fibroblast growth factor-2 counteracts the effect of ciliary neurotrophic factor on spontaneous differentiation in adult hippocampal progenitor cells

Abstract

Neural stem/progenitor cells (NSCs) can spontaneously differentiate into neurons and glial cells in the absence of mitogen fibroblast growth factor-2 (FGF-2) or epidermal growth factor (EGF) in medium and the spontaneous differentiation of NSCs is mediated partially by endogenous ciliary neurotrophic factor (CNTF). This study examined the relationship of FGF-2 and CNTF in the spontaneous differentiation of adult hippocampal progenitor cells (AHPs). AHPs were cultured in the medium containing different concentration of FGF-2 (1–100 ng/mL). Western blotting and immunofluorescence staining were applied to detect the expression of the astrocytic marker GFAP, the neuronal marker Tuj1, the oligodendrocytic marker CNPase and, Nestin, the marker of AHPs. The expression of endogenous CNTF in AHPs at early (passage 4) and late stage (passage 22) was also measured by Western blotting. The results showed that FGF-2 increased the expression of Nestin, dramatically inhibited the expression of GFAP and Tuj1 and slightly suppressed the expression of CNPase. FGF-2 down-regulated the expression of endogenous CNTF in AHPs at both early (passage 4) and late stage (passage 22). These results suggested that FGF-2 could inhibit the spontaneous differentiation of cultured AHPs by negatively regulating the expression of endogenous CNTF in AHPs.

Keywords

spontaneous differentiation / neural progenitor cells / basic fibroblast growth factor / neurogenesis

References

[1] ReynoldsB.A., WeissS.. Generation of neurons and astrocytes from isolated cells of the adult mammalian central nervous system. Science, 1992, 255(5052): 1707-1710

[2] ReynoldsB.A., TetzlaffW., WeissS.. A multipotent EGF-responsive striatal embryonic progenitor cell produces neurons and astrocytes. J Neurosci, 1992, 12(11): 4565-4574

[3] RichardsL.J., KilpatrickT.J., BartlettP.F.. De novo generation of neuronal cells from the adult mouse brain. Proc Natl Acad Sci USA, 1992, 89(18): 8591-8595

[4] GouldE., CameronH.A., DanielsD.C., et al.. Adrenal hormones suppress cell division in the adult rat dentate gyrus. J Neurosci, 1992, 12(9): 3642-3650

[5] CorottoF.S., HenegarJ.A., MaruniakJ.A.. Neurogenesis persists in the subependymal layer of the adult mouse brain. Neurosci Lett, 1993, 149(2): 111-114

[6] LuskinM.B.. Restricted proliferation and migration of postnatally generated neurons derived from the forebrain subventricular zone. Neuron, 1993, 11(1): 173-189

[7] KornackD.R., RakicP.. The generation, migration, and differentiation of olfactory neurons in the adult primate brain. Proc Natl Acad Sci USA, 2001, 98(8): 4752-4757

[8] van PraagH., SchinderA.F., ChristieB.R., et al.. Functional neurogenesis in the adult hippocampus. Nature, 2002, 415(6875): 1030-1034

[9] ReynoldsB.A., WeissS.. Clonal and population analyses demonstrate that an EGF-responsive mammalian embryonic CNS precursor is a stem cell. Dev Biol, 1996, 175(1): 1-13

[10] KilpatrickT.J., BartlettP.F.. Cloning and growth of multipotential neural precursors: requirements for proliferation and differentiation. Neuron, 1993, 10(2): 255-265

[11] WeissS., DunneC., HewsonJ., et al.. Multipotent CNS stem cells are present in the adult mammalian spinal cord and ventricular neuroaxis. J Neurosci, 1996, 16(23): 7599-7609

[12] WeissS., van der KooyD.. CNS stem cells: where’s the biology (a.k.a. beef)?. J Neurobiol, 1998, 36(2): 307-314

[13] DuZ.W., ZhangS.C.. Neural differentiation from embryonic stem cells: which way?. Stem Cells Dev, 2004, 13(4): 372-381

[14] Wislet-GendebienS., BruyereF., HansG., et al.. Nestin-positive mesenchymal stem cells favour the astroglial lineage in neural progenitors and stem cells by releasing active BMP4. BMC Neurosci, 2004, 5: 33

[15] HermansonO., JepsenK., RosenfeldM.G.. N-CoR controls differentiation of neural stem cells into astrocytes. Nature, 2002, 419(6910): 934-939

[16] AngelastroJ.M., MasonJ.L., IgnatovaT.N., et al.. Downregulation of activating transcription factor 5 is required for differentiation of neural progenitor cells into astrocytes. J Neurosci, 2005, 25(15): 3889-3899

[17] YoneyamaM., FukuiM., NakamichiN., et al.. Activation of GABA(A) receptors facilitates astroglial differentiation induced by ciliary neurotrophic factor in neural progenitors isolated from fetal rat brain. J Neurochem, 2007, 100(6): 1667-1679

[18] LouisJ.C., MagalE., TakayamaS., et al.. CNTF protection of oligodendrocytes against natural and tumor necrosis factor-induced death. Science, 1993, 259(5095): 689-692

[19] ChenH., TungY.C., LiB., et al.. Trophic factors counteract elevated FGF-2-induced inhibition of adult neurogenesis. Neurobiol Aging, 2007, 28(8): 1148-1162

[20] KangS.S., KeaseyM.P., ArnoldS.A., et al.. Endogenous CNTF mediates stroke-induced adult CNS neurogenesis in mice. Neurobiol Dis, 2012, 31(49): 68-78

[21] VescoviA.L., ReynoldsB.A., FraserD.D., et al.. bFGF regulates the proliferative fate of unipotent (neuronal) and bipotent (neuronal/astroglial) EGF-generated CNS progenitor cells. Neuron, 1993, 11(5): 951-966

[22] KilpatrickT.J., BartlettP.F.. Cloned multipotential precursors from the mouse cerebrum require FGF-2, whereas glial restricted precursors are stimulated with either FGF-2 or EGF. J Neurosci, 1995, 15(5Pt1): 3653-3661

[23] GrittiA., Frolichsthal-SchoellerP., GalliR., et al.. Epidermal and fibroblast growth factors behave as mitogenic regulators for a single multipotent stem cell-like population from the subventricular region of the adult mouse forebrain. J Neurosci, 1999, 19(9): 3287-3297

[24] TropepeV., SibiliaM., CirunaB.G., et al.. Distinct neural stem cells proliferate in response to EGF and FGF in the developing mouse telencephalon. Dev Biol, 1999, 208(1): 166-188

[25] BensadounA., WeinsteinD.. Assay of proteins in the presence of interfering materials. Anal Biochem, 1976, 70: 241-250

[26] GalliR., GrittiA., BonfantiL., et al.. Neural stem cells: an overview. Circ Res, 2003, 92(6): 598-608

[27] KuhnH.G., WinklerJ., KempermannG., et al.. Epidermal growth factor and fibroblast growth factor-2 have different effects on neural progenitors in the adult rat brain. J Neurosci, 1997, 17(15): 5820-5829

[28] MaricD., MaricI., ChangY.H., et al.. Prospective cell sorting of embryonic rat neural stem cells and neuronal and glial progenitors reveals selective effects of basic fibroblast growth factor and epidermal growth factor on self-renewal and differentiation. J Neurosci, 2003, 23(1): 240-251

[29] DeleyrolleL., Marchal-VictorionS., DromardC., et al.. Exogenous and fibroblast growth factor 2/epidermal growth factor-regulated endogenous cytokines regulate neural precursor cell growth and differentiation. Stem Cells, 2006, 24(3): 748-762

[30] CummingsB.J., SuJ.H., CotmanC.W.. Neuritic involvement within FGF-2 immunopositive plaques of Alzheimer disease. Exp Neuro, 1993, 124: 315-125