Understanding the mechanisms regulating retinal stem cell (RSC) activity is fundamental for future stem cell-based therapeutic purposes. By combining gain and loss of function approaches, we addressed whether Notch signaling may play a selective role in retinal stem versus retinal progenitor cells in both developing and adult eyes. Inhibition of either Notch or fibroblast growth factor signaling reduced proliferation of retinal stem and retinal progenitor cells, and inhibited RSC self-renewal. Conversely, exogenous Delta-like 3 and direct intrinsic Notch activation stimulated expansionary symmetric divisions in adult RSCs with the concomitant upregulation of Hes5. Knocking down Hes5 expression specifically decreased the numbers, but not the diameters, of adult RSC primary spheres, indicating that HES5 is the downstream effector of Notch receptor in controlling adult RSC proliferation. In addition, constitutive Notch activation induced retinal stem-like asymmetric self-renewal properties, with no expansion (no symmetrical division) in perinatal neural retina progenitor cells. These findings highlight central roles of Notch signaling activity in regulating the modes of division of retinal stem and retinal progenitor cells.
Get full access to this article
View all access options for this article.
References
1.
OhlsteinB, KaiT, DecottoE and SpradlingA. (2004). The stem cell niche: theme and variations. Curr Opin Cell Biol, 16:693–699.
2.
WalkerMR, PatelKK and StappenbeckTS. (2009). The stem cell niche. J Pathol, 217:169–180.
3.
JohnsPR. (1977). Growth of the adult goldfish eye. III. Source of the new retinal cells. J Comp Neurol, 176:343–357.
4.
WettsR, SerbedzijaGN and FraserSE. (1989). Cell lineage analysis reveals multipotent precursors in the ciliary margin of the frog retina. Dev Biol, 136:254–263.
5.
OttesonDC and HitchcockPF. (2003). Stem cells in the teleost retina: persistent neurogenesis and injury-induced regeneration. Vis Res, 43:927–936.
6.
BernardosRL, BarthelLK, MeyersJR and RaymondPA. (2007). Late-stage neuronal progenitors in the retina are radial Muller glia that function as retinal stem cells. J Neurosci, 27:7028–7040.
7.
RaymondPA, BarthelLK, BernardosRL and PerkowskiJJ. (2006). Molecular characterization of retinal stem cells and their niches in adult zebrafish. BMC Dev Biol, 6:36
8.
FischerAJ and BonginiR. (2010). Turning Muller glia into neural progenitors in the retina. Mol Neurobiol, 42:199–209.
9.
PerronM and HarrisWA. (2000). Retinal stem cells in vertebrates. Bioessays, 22:685–688.
10.
RehTA and FischerAJ. (2006). Retinal stem cells. Methods Enzymol, 419:52–73.
11.
MoshiriA, CloseJ and RehTA. (2004). Retinal stem cells and regeneration. Int J Dev Biol, 48:1003–1014.
12.
FangY, ChoKS, TchedreK, LeeSW, GuoC, KinouchiH, FriedS, SunX and ChenDF. (2013). Ephrin-A3 suppresses Wnt signaling to control retinal stem cell potency. Stem Cells, 31:349–359.
13.
ColesBL, HorsfordDJ, McInnesRR and van der KooyD. (2006). Loss of retinal progenitor cells leads to an increase in the retinal stem cell population in vivo. Eur J Neurosci, 23:75–82.
14.
MoshiriA and RehTA. (2004). Persistent progenitors at the retinal margin of ptc+/− mice. J Neurosci, 24:229–237.
15.
DucournauY, BoscherC, AdelmanRA, GuillaubeyC, Schmidt-MorandD, MosnierJF and DucournauD. (2012). Proliferation of the ciliary epithelium with retinal neuronal and photoreceptor cell differentiation in human eyes with retinal detachment and proliferative vitreoretinopathy. Graefes Arch Clin Exp Ophthalmol, 250:409–423.
16.
GualdoniS, BaronM, LakowskiJ, DecembriniS, SmithAJ, PearsonRA, AliRR and SowdenJC. (2010). Adult ciliary epithelial cells, previously identified as retinal stem cells with potential for retinal repair, fail to differentiate into new rod photoreceptors. Stem Cells, 28:1048–1059.
17.
CiceroSA, JohnsonD, ReyntjensS, FraseS, ConnellS, ChowLM, BakerSJ, SorrentinoBP and DyerMA. (2009). Cells previously identified as retinal stem cells are pigmented ciliary epithelial cells. Proc Natl Acad Sci U S A, 106:6685–6690.
18.
Guduric-FuchsJ, ChenW, PriceH, ArcherDB and CogliatiT. (2011). RPE and neuronal differentiation of allotransplantated porcine ciliary epithelium-derived cells. Mol Vis, 17:2580–2595.
19.
MoeMC, KolbergRS, SandbergC, Vik-MoE, OlstornH, VargheseM, LangmoenIA and NicolaissenB. (2009). A comparison of epithelial and neural properties in progenitor cells derived from the adult human ciliary body and brain. Exp Eye Res, 88:30–38.
20.
Lord-GrignonJ, AbdouhM and BernierG. (2006). Identification of genes expressed in retinal progenitor/stem cell colonies isolated from the ocular ciliary body of adult mice. Gene Expr Patterns, 6:992–999.
21.
InoueY, YanagiY, TamakiY, UchidaS, KawaseY, AraieM and OkochiH. (2005). Clonogenic analysis of ciliary epithelial derived retinal progenitor cells in rabbits. Exp Eye Res, 81:437–445.
22.
ColesBL, AngenieuxB, InoueT, Del Rio-TsonisK, SpenceJR, McInnesRR, ArsenijevicY and van der KooyD. (2004). Facile isolation and the characterization of human retinal stem cells. Proc Natl Acad Sci U S A, 101:15772–15777.
23.
TropepeV, ColesBL, ChiassonBJ, HorsfordDJ, EliaAJ, McInnesRR and van der KooyD. (2000). Retinal stem cells in the adult mammalian eye. Science, 287:2032–2036.
24.
BalliosBG, ClarkeL, ColesBLK, ShoichetMS and Van Der KooyD. (2012). The adult retinal stem cell is a rare cell in the ciliary epithelium whose progeny can differentiate into photoreceptors. 237–246.
25.
BeebeDC. (1986). Development of the ciliary body: a brief review. Trans Ophthalmol Soc U K, 105 (Pt 2):123–130.
26.
YoungRW. (1985). Cell proliferation during postnatal development of the retina in the mouse. Brain Res, 353:229–239.
27.
AguirreA, RubioME and GalloV. (2010). Notch and EGFR pathway interaction regulates neural stem cell number and self-renewal. Nature, 467:323–327.
28.
CepkoCL. (1999). The roles of intrinsic and extrinsic cues and bHLH genes in the determination of retinal cell fates. Curr Opin Neurobiol, 9:37–46.
29.
HitoshiS, AlexsonT, TropepeV, DonovielD, EliaAJ, NyeJS, ConlonRA, MakTW, BernsteinA and van der KooyD. (2002). Notch pathway molecules are essential for the maintenance, but not the generation, of mammalian neural stem cells. Genes Dev, 16:846–858.
30.
LiveseyFJ and CepkoCL. (2001). Vertebrate neural cell-fate determination: lessons from the retina. Nat Rev, 2:109–118.
31.
LouviA and Artavanis-TsakonasS. (2006). Notch signalling in vertebrate neural development. Nat Rev, 7:93–102.
32.
MizutaniK, YoonK, DangL, TokunagaA and GaianoN. (2007). Differential Notch signalling distinguishes neural stem cells from intermediate progenitors. Nature, 449:351–355.
33.
YunK, FischmanS, JohnsonJ, Hrabe de AngelisM, WeinmasterG and RubensteinJL. (2002). Modulation of the notch signaling by Mash1 and Dlx1/2 regulates sequential specification and differentiation of progenitor cell types in the subcortical telencephalon. Development, 129:5029–5040.
34.
ChiassonBJ, TropepeV, MorsheadCM and van der KooyD. (1999). Adult mammalian forebrain ependymal and subependymal cells demonstrate proliferative potential, but only subependymal cells have neural stem cell characteristics. J Neurosci, 19:4462–4471.
Artavanis-TsakonasS, RandMD and LakeRJ. (1999). Notch signaling: cell fate control and signal integration in development. Science, 284:770–776.
37.
BaoZZ and CepkoCL. (1997). The expression and function of Notch pathway genes in the developing rat eye. J Neurosci, 17:1425–1434.
38.
DorskyRI, ChangWS, RapaportDH and HarrisWA. (1997). Regulation of neuronal diversity in the Xenopus retina by Delta signalling. Nature, 385:67–70.
39.
DorskyRI, RapaportDH and HarrisWA. (1995). Xotch inhibits cell differentiation in the Xenopus retina. Neuron, 14:487–496.
40.
HenriqueD, HirsingerE, AdamJ, Le RouxI, PourquieO, Ish-HorowiczD and LewisJ. (1997). Maintenance of neuroepithelial progenitor cells by Delta-Notch signalling in the embryonic chick retina. Curr Biol, 7:661–670.
41.
JadhavAP, ChoSH and CepkoCL. (2006). Notch activity permits retinal cells to progress through multiple progenitor states and acquire a stem cell property. Proc Natl Acad Sci U S A, 103:18998–19003.
42.
AlexsonTO, HitoshiS, ColesBL, BernsteinA and van der KooyD. (2006). Notch signaling is required to maintain all neural stem cell populations—irrespective of spatial or temporal niche. Dev Neurosci, 28:34–48.
43.
HojoM, OhtsukaT, HashimotoN, GradwohlG, GuillemotF and KageyamaR. (2000). Glial cell fate specification modulated by the bHLH gene Hes5 in mouse retina. Development, 127:2515–2522.
44.
TurnerDL and CepkoCL. (1987). A common progenitor for neurons and glia persists in rat retina late in development. Nature, 328:131–136.
45.
NelsonBR, HartmanBH, GeorgiSA, LanMS and RehTA. (2007). Transient inactivation of Notch signaling synchronizes differentiation of neural progenitor cells. Dev Biol, 304:479–498.
46.
DavisDM and DyerMA. (2010). Retinal progenitor cells, differentiation, and barriers to cell cycle reentry. Curr Top Dev Biol, 93:175–188.
JadhavAP, MasonHA and CepkoCL. (2006). Notch 1 inhibits photoreceptor production in the developing mammalian retina. Development, 133:913–923.
49.
FurukawaT, MukherjeeS, BaoZZ, MorrowEM and CepkoCL. (2000). rax, Hes1, and notch1 promote the formation of Muller glia by postnatal retinal progenitor cells. Neuron, 26:383–394.
50.
DuncanAW, RattisFM, DiMascioLN, CongdonKL, PazianosG, ZhaoC, YoonK, CookJM, WillertK, GaianoN and ReyaT. (2005). Integration of Notch and Wnt signaling in hematopoietic stem cell maintenance. Nat Immunol, 6:314–322.
51.
YasugiT, SugieA, UmetsuD and TabataT. (2010). Coordinated sequential action of EGFR and Notch signaling pathways regulates proneural wave progression in the Drosophila optic lobe. Development, 137:3193–3203.
52.
EggerB, GoldKS and BrandAH. (2010). Notch regulates the switch from symmetric to asymmetric neural stem cell division in the Drosophila optic lobe. Development, 137:2981–2987.
53.
ShimojoH, OhtsukaT and KageyamaR. (2008). Oscillations in notch signaling regulate maintenance of neural progenitors. Neuron, 58:52–64.
54.
GeorgiSA and RehTA. (2011). Dicer is required for the maintenance of Notch signaling and gliogenic competence during mouse retinal development. Dev Neurobiol, 71:1153–1169.
55.
ImayoshiI, SakamotoM, YamaguchiM, MoriK and KageyamaR. (2010). Essential roles of Notch signaling in maintenance of neural stem cells in developing and adult brains. J Neurosci, 30:3489–3498.
56.
LugertS, BasakO, KnucklesP, HausslerU, FabelK, GotzM, HaasCA, KempermannG, TaylorV and GiachinoC. (2010). Quiescent and active hippocampal neural stem cells with distinct morphologies respond selectively to physiological and pathological stimuli and aging. Cell Stem Cell, 6:445–456.
57.
SanalkumarR, IndulekhaCL, DivyaTS, DivyaMS, AntoRJ, VinodB, VidyanandS, JagathaB, VenugopalS and JamesJ. (2010). ATF2 maintains a subset of neural progenitors through CBF1/Notch independent Hes-1 expression and synergistically activates the expression of Hes-1 in Notch-dependent neural progenitors. J Neurochem, 113:807–818.
58.
WallDS, MearsAJ, McNeillB, MazerolleC, ThurigS, WangY, KageyamaR and WallaceVA. (2009). Progenitor cell proliferation in the retina is dependent on Notch-independent Sonic hedgehog/Hes1 activity. J Cell Biol, 184:101–112.
59.
SprinzakD, LakhanpalA, LebonL, SantatLA, FontesME, AndersonGA, Garcia-OjalvoJ and ElowitzMB. (2010). Cis-interactions between Notch and Delta generate mutually exclusive signalling states. Nature, 465:86–90.
60.
NelsonBR, HartmanBH, RayCA, HayashiT, Bermingham-McDonoghO and RehTA. (2009). Acheate-scute like 1 (Ascl1) is required for normal delta-like (Dll) gene expression and notch signaling during retinal development. Dev Dyn, 238:2163–2178.
61.
NelsonBR and RehTA. (2008). Relationship between Delta-like and proneural bHLH genes during chick retinal development. Dev Dyn, 237:1565–1580.
Supplementary Material
Please find the following supplemental material available below.
For Open Access articles published under a Creative Commons License, all supplemental material carries the same license as the article it is associated with.
For non-Open Access articles published, all supplemental material carries a non-exclusive license, and permission requests for re-use of supplemental material or any part of supplemental material shall be sent directly to the copyright owner as specified in the copyright notice associated with the article.
