Current immunosuppressive treatments for central nervous system demyelinating diseases fail to prevent long-term motor and cognitive decline in patients. Excitingly, glial cell transplantation arises as a promising complementary strategy to challenge oligodendrocytes loss occurring in myelination disorders. A potential source of new oligodendrocytes is the subventricular zone (SVZ) pool of multipotent neural stem cells. However, this approach has been handicapped by the lack of functional methods for identification and pharmacological analysis of differentiating oligodendrocytes, prior to transplantation. In this study, we questioned whether SVZ-derived oligodendrocytes could be functionally discriminated due to intracellular calcium level ([Ca2+]i) variations following KCl, histamine, and thrombin stimulations. Previously, we have shown that SVZ-derived neurons and immature cells can be discriminated on the basis of their selective [Ca2+]i rise upon KCl and histamine stimulation, respectively. Herein, we demonstrate that O4+ and proteolipid protein-positive (PLP+) oligodendrocytes do not respond to these stimuli, but display a robust [Ca2+]i rise following thrombin stimulation, whereas other cell types are thrombin-insensitive. Thrombin-induced Ca2+ increase in oligodendrocytes is mediated by protease-activated receptor-1 (PAR-1) activation and downstream signaling through Gq/11 and phospholipase C (PLC), resulting in Ca2+ recruitment from intracellular compartments. This method allows the analysis of functional properties of oligodendrocytes in living SVZ cultures, which is of major interest for the development of effective grafting strategies in the demyelinated brain. Additionally, it opens new perspectives for the search of new pro-oligodendrogenic factors to be used prior grafting.
Get full access to this article
View all access options for this article.
References
1.
FranklinRJ, Ffrench-ConstantC. Remyelination in the CNS: From biology to therapy. Nat Rev Neurosci, 2008; 9:839–855.
2.
LevisonSW, GoldmanJE. Both oligodendrocytes and astrocytes develop from progenitors in the subventricular zone of postnatal rat forebrain. Neuron, 1993; 10:201–212.
3.
MennB, Garcia-VerdugoJM, YaschineC, Gonzalez-PerezO, RowitchD, Alvarez-BuyllaA. Origin of oligodendrocytes in the subventricular zone of the adult brain. J Neurosci, 2006; 26:7907–7918.
4.
CayreM, BancilaM, VirardI, BorgesA, DurbecP. Migrating and myelinating potential of subventricular zone neural progenitor cells in white matter tracts of the adult rodent brain. Mol Cell Neurosci, 2006; 31:748–758.
5.
SmithPM, BlakemoreWF. Porcine neural progenitors require commitment to the oligodendrocyte lineage prior to transplantation in order to achieve significant remyelination of demyelinated lesions in the adult CNS. Eur J Neurosci, 2000; 12:2414–2424.
6.
KeirsteadHS, Ben-HurT, RogisterB, O'LearyMT, Dubois-DalcqM, BlakemoreWF. Polysialylated neural cell adhesion molecule-positive CNS precursors generate both oligodendrocytes and schwann cells to remyelinate the CNS after transplantation. J Neurosci, 1999; 19:7529–7536.
7.
AkiyamaY, HonmouO, KatoT, UedeT, HashiK, KocsisJD. Transplantation of clonal neural precursor cells derived from adult human brain establishes functional peripheral myelin in the rat spinal cord. Exp Neurol, 2001; 167:27–39.
8.
AgasseF, BernardinoL, SilvaB, FerreiraR, GradeS, MalvaJO. Response to histamine allows the functional identification of neuronal progenitors, neurons, astrocytes, and immature cells in subventricular zone cell cultures. Rejuvenation Res, 2008; 11:187–200.
BernardinoL, AgasseF, SilvaB, FerreiraR, GradeS, MalvaJO. Tumor necrosis factor-alpha modulates survival, proliferation, and neuronal differentiation in neonatal subventricular zone cell cultures. Stem Cells, 2008; 26:2361–2371.
11.
WangY, Richter-LandsbergC, ReiserG. Expression of protease-activated receptors (PARs) in OLN-93 oligodendroglial cells and mechanism of PAR-1-induced calcium signaling. Neuroscience, 2004; 126:69–82.
12.
SilvaCG, PorciúnculaLO, CanasPM, OliveiraCR, CunhaRA. Blockade of adenosine A(2A) receptors prevents staurosporine-induced apoptosis of rat hippocampal neurons. Neurobiol Dis, 2007; 27:182–189.
13.
MatosM, AugustoE, OliveiraCR, AgostinhoP. Amyloid-beta peptide decreases glutamate uptake in cultured astrocytes: involvement of oxidative stress and mitogen-activated protein kinase cascades. Neuroscience, 2008; 156:898–910.
14.
CoughlinSR. How the protease thrombin talks to cells. Proc Natl Acad Sci USA, 1999; 96:11023–11027.
15.
JacquesSL, KuliopulosA. Protease-activated receptor-4 uses dual prolines and an anionic retention motif for thrombin recognition and cleavage. Biochem J, 2003; 376:733–740.
16.
Nakanishi-MatsuiM, ZhengYW, SulcinerDJ, WeissEJ, LudemanMJ, CoughlinSR. PAR3 is a cofactor for PAR4 activation by thrombin. Nature, 2000; 404:609–613.
17.
AhnHS, FosterC, BoykowG, StamfordA, MannaM, GrazianoM. Inhibition of cellular action of thrombin by n3-cyclopropyl-7-[4-(1-methylethyl) phenyl] methyl}-7h-pyrrolo [3, 2-f] quinazoline-1, 3-diamine (SCH 79797), a nonpeptide thrombin receptor antagonist. Biochem Pharmacol, 2000; 60:1425–1434.
18.
FernándezM, ParadisiM, Del VecchioG, GiardinoL, CalzàL. Thyroid hormone induces glial lineage of primary neurospheres derived from non-pathological and pathological rat brain: Implications for remyelination-enhancing therapies. Int J Dev Neurosci, 2009; 27:769–778.
19.
FrancoPG, SilvestroffL, SotoEF, PasquiniJM. Thyroid hormones promote differentiation of oligodendrocyte progenitor cells and improve remyelination after cuprizone-induced demyelination. Exp Neurol, 2008; 212:458–467.
20.
JoheKK, HazelTG, MullerT, Dugich-DjordjevicMM, McKayRD. Single factors direct the differentiation of stem cells from the fetal and adult central nervous system. Genes Dev, 1996; 10:3129–3140.
21.
BaumannN, Pham-DinhD. Biology of oligodendrocyte and myelin in the mammalian central nervous system. Physiol Rev, 2001; 81:871–927.
22.
AguirreAA, ChittajalluR, BelachewS, GalloV. NG2-expressing cells in the subventricular zone are type C-like cells and contribute to interneuron generation in the postnatal hippocampus. J Cell Biol, 2004; 165:575–589.
23.
CoughlinSR. Thrombin signalling and protease-activated receptors. Nature, 2000; 407:258–264.
McLaughlinJN, ShenL, HolinstatM, BrooksJD, DiBenedettoE, HammHE. Functional selectivity of G protein signaling by agonist peptides and thrombin for the protease-activated receptor-1. J Biol Chem, 2005; 280:25048–25059.
26.
HainsMD, WingMR, MaddiletiS, SiderovskiDP, HardenTK. Galpha12/13- and rho-dependent activation of phospholipase c-epsilon by lysophosphatidic acid and thrombin receptors. Mol Pharmacol, 2006; 69:2068–2075.
27.
WhittemoreSR, MorassuttiDJ, WaltersWM, LiuRH, MagnusonDS. Mitogen and substrate differentially affect the lineage restriction of adult rat subventricular zone neural precursor cell populations. Exp Cell Res, 1999; 252:75–95.
28.
GrittiA, ParatiEA, CovaL, FrolichsthalP, GalliR, WankeE, FaravelliL, MorassuttiDJ, RoisenF, NickelDD, VescoviAL. Multipotential stem cells from the adult mouse brain proliferate and self-renew in response to basic fibroblast growth factor. J Neurosci, 1996; 16:1091–1100.
29.
BarresBA, LazarMA, RaffMC. A novel role for thyroid hormone, glucocorticoids and retinoic acid in timing oligodendrocyte development. Development, 1994; 120:1097–1108.
30.
ZhangP, ChebathJ, LonaiP, RevelM. Enhancement of oligodendrocyte differentiation from murine embryonic stem cells by an activator of gp130 signaling. Stem Cells, 2004; 22:344–354.
31.
KangSM, ChoMS, SeoH, YoonCJ, OhSK, ChoiYM, KimDW. Efficient induction of oligodendrocytes from human embryonic stem cells. Stem Cells, 2007; 25:419–424.
32.
MurrayK, Dubois-DalcqM. Emergence of oligodendrocytes from human neural spheres. J Neurosci Res, 1997; 50:146–156.
33.
RavinR, HoeppnerDJ, MunnoDM, CarmelL, SullivanJ, LevittDL, MillerJL, AthaideC, PanchisionDM, McKayRD. Potency and fate specification in CNS stem cell populations in vitro. Cell Stem Cell, 2008; 3:670–680.
34.
JinK, MaoXO, SunY, XieL, GreenbergDA. Stem cell factor stimulates neurogenesis in vitro and in vivo. J Clin Invest, 2002; 110:311–319.
35.
LaywellED, RakicP, KukekovVG, HollandEC, SteindlerDA. Identification of a multipotent astrocytic stem cell in the immature and adult mouse brain. Proc Natl Acad Sci USA, 2000; 97:13883–13888.
36.
Rodríguez-PeñaA. Oligodendrocyte development and thyroid hormone. J Neurobiol, 1999; 40:497–512.
37.
BernalJ. Thyroid hormones and brain development. Vitam Horm, 2005; 71:95–122.
38.
BaasD, LegrandC, SamarutJ, FlamantF. Persistence of oligodendrocyte precursor cells and altered myelination in optic nerve associated to retina degeneration in mice devoid of all thyroid hormone receptors. Proc Natl Acad Sci USA, 2002; 99:2907–2911.
39.
ShavitE, BeilinO, KorczynAD, SylantievC, AronovichR, DroryVE, GurwitzD, HorreshI, Bar-ShavitR, PelesE, ChapmanJ. Thrombin receptor PAR-1 on myelin at the node of Ranvier: a new anatomy and physiology of conduction block. Brain, 2008; 131:1113–1122.
40.
SmirnovaIV, CitronBA, ArnoldPM, FestoffBW. Neuroprotective signal transduction in model motor neurons exposed to thrombin: G-protein modulation effects on neurite outgrowth, Ca2+ mobilization, and apoptosis. J. Neurobiol, 2001; 48:87–100.
41.
OlianasMC, DedoniS, OnaliP. Proteinase-activated receptors 1 and 2 in rat olfactory system: Layer-specific regulation of multiple signaling pathways in the main olfactory bulb and induction of neurite retraction in olfactory sensory neurons. Neuroscience, 2007; 146:1289–1301.
42.
GorbachevaLR, StorozhevykhTP, PinelisVG, IshiwataS, StrukovaSM. Modulation of hippocampal neuron survival by thrombin and factor Xa. Biochemistry (Moscow), 2006; 71:1082–1089.
43.
YangY, AkiyamaH, FentonJW2nd, BrewerGJ. Thrombin receptor on rat primary hippocampal neurons: Coupled calcium and cAMP responses. Brain Res, 1997; 761:11–18.
44.
WangH, UblJJ, ReiserG. Four subtypes of protease-activated receptors, co-expressed in rat astrocytes, evoke different physiological signaling. Glia, 2002; 37:53–63.
45.
SorensenSD, NicoleO, PeavyRD, MontoyaLM, LeeCJ, MurphyTJ, TraynelisSF, HeplerJR. Common signaling pathways link activation of murine PAR-1, LPA, and S1P receptors to proliferation of astrocytes. Mol Pharmacol, 2003; 64:1199–1209.
