Journal of Molecular Neuroscience

, Volume 42, Issue 1, pp 28–34 | Cite as

The Expression of Neuroepithelial Cell Fate Determinants in Rat Spinal Cord Development



Lineage specification is tightly regulated by a unique combination of extrinsic and intrinsic cues, but exactly how these cues coordinate the timing and position of cell differentiation during spinal cord development needs further investigation. Notch signaling has major roles in lineage specification. Recent evidence also indicates that the combination of transcription factors of the basic helix-loop-helix (Hes3, Hes5) and homeodomain (Pax6) families establish molecular codes that determine both the timing and position of neurons and glia. The precise expression patterns of these genes in vivo in the developing spinal cord from E13 to E18 are not fully known. In this study, the spatial and temporal expression patterns of these genes have been investigated. RT-PCR studies reveal the differential expression of these genes. The dynamic changes detected in the expression of these molecules have an important role in spinal cord cell lineage specification. Moreover, this study clarifies their in vivo expression during spinal cord development, and the expression patterns observed shed light on the generation of the rostro-caudal gradient of development. By understanding how neural stem cells are regulated in spinal cord development in vivo, we may gain insight of relevance to cell replacement strategies to treat spinal cord injuries.


Pax6 Notch1 Hes3 Hes5 Cell fate Spinal cord 



This work was funded by the Health Research Board Ireland. Many thanks to Dr. Siobhain O’Mahony, Dr. Gerard Clarke, and Bereniece Riedewald for technical advice and to Bernadette O’Donovan for some laboratory assistance.


  1. Akazawa C, Sasai Y, Nakanishi S, Kageyama R (1992) Molecular characterization of a rat negative regulator with a basic helix-loop-helix structure predominantly expressed in the developing nervous system. J Biol Chem 267:21879–21885PubMedGoogle Scholar
  2. Alvarez-Buylla A, García-Verdugo JM, Tramontin AD (2001) A unified hypothesis on the lineage of neural stem cells. Nat Rev Neurosci 2:287–293CrossRefPubMedGoogle Scholar
  3. Artavanis-Tsakonas S, Rand MD, Lake RJ (1999) Notch signaling: cell fate control and signal integration in development. Science 284:770–776CrossRefPubMedGoogle Scholar
  4. Bao ZZ, Cepko CL (1997) The expression and function of notch pathway genes in the developing rat eye. J Neurosci 17(4):1425–1434PubMedGoogle Scholar
  5. Barry D, McDermott K (2005) Differentiation of radial glia from radial precursor cells and transformation into astrocytes in the developing rat spinal cord. Glia 50:187–197CrossRefPubMedGoogle Scholar
  6. Bel-Vialar S, Medevielle F, Pituello F (2007) The on/off of Pax6 controls the tempo of neuronal differentiation in the developing spinal cord. Dev Biol 305:659–673CrossRefPubMedGoogle Scholar
  7. Chambers CB, Peng Y, Nguyen H, Gaiano N, Fishell G, Nye JS (2001) Spatiotemporal selectivity of response to Notch1 signals in mammalian forebrain precursors. Development 128:689–702PubMedGoogle Scholar
  8. Choi BH, Lapham LW (1978) Radial glia in the human fetal cerebrum: a combined Golgi, immunofluorescent and electron microscopic study. Brain Res 148(2):295–311CrossRefPubMedGoogle Scholar
  9. de Azevedo LC, Fallet C, Moura-Neto V, Daumas-Duport C, Hedin-Pereira C, Lent R (2003) Cortical radial glial cells in human fetuses: depth-correlated transformation into astrocytes. J Neurobiol 55(3):288–298CrossRefGoogle Scholar
  10. de la Pompa JL, Wakeham A, Correia KM, Samper E, Brown S, Aguilera RJ, Nakano T, Honjo T, Mak TW, Rossant J, Conlon RA (1997) Conservation of the Notch signalling pathway in mammalian neurogenesis. Development 124:1139–1148PubMedGoogle Scholar
  11. Ericson J, Rashbass P, Schedl A, Brenner-Morton S, Kawakami A, van Heyningen V, Jessell TM, Briscoe J (1997) Pax6 controls progenitor cell identity and neuronal fate in response to graded Shh signalling. Cell 90:169–180CrossRefPubMedGoogle Scholar
  12. Frisén J, Lendahl U (2001) Oh no, Notch again! Bioessays 23:3–7CrossRefPubMedGoogle Scholar
  13. Gaiano N, Nye JS, Fishell G (2000) Radial glial identity is promoted by Notch1 signaling in the murine forebrain. Neurona 26:395–404CrossRefGoogle Scholar
  14. Gavalas A, Krumlauf R (2000) Retinoid signalling and hindbrain patterning. Curr Opin Genet Dev 10(4):380–386CrossRefPubMedGoogle Scholar
  15. Hatakeyama J, Bessho Y, Katoh K, Ookawara S, Fujioka M, Guillemot F, Kageyama R (2004) Hes genes regulate size, shape and histogenesis of the nervous system by control of the timing of neural stem cell differentiation. Development 131:5539–5550CrossRefPubMedGoogle Scholar
  16. Henrique D, Hirsinger E, Adam J, Le Roux I, Pourquié O, Ish-Horowicz D, Lewis J (1997) Maintenance of neuroepithelial progenitor cells by Delta-Notch signaling in the embryonic chick retina. Curr Biol 7:661–670CrossRefPubMedGoogle Scholar
  17. Higuchi M, Kiyama H, Hayakawa T, Hamada Y, Tsujimoto Y (1995) Differential expression of Notch1 and Notch2 in developing and adult mouse brain. Mol Brain Res 29:263–272CrossRefPubMedGoogle Scholar
  18. Hirata H, Ohtsuka T, Bessho Y, Kageyama R (2000) Generation of structurally and functionally distinct factors from the basic helix-loop-helix gene Hes3 by alternative first exons. J Biol Chem 275:19083–19089CrossRefPubMedGoogle Scholar
  19. Hitoshi S, Alexson T, Tropepe V, Donoviel D, Elia AJ, Nye JS, Conlon RA, Mak TW, Bernstein A, van der Kooy D (2002) Notch pathway molecules are essential for the maintenance, but not the generation of mammalian neural stem cells. Gene Dev 16:846–858CrossRefPubMedGoogle Scholar
  20. Hojo M, Ohtsuka T, Hashimoto N, Gradwohl G, Guillemot F, Kageyama R (2000) Glial cell fate specification modulated by the bHLH gene Hes5 in mouse retina. Development 127:2515–2522PubMedGoogle Scholar
  21. Ishibashi M, Moriyoshi K, Sasail Y, Shiota K, Nakanishi S, Kageyama R (1994) Persistent expression of helix-loop-helix factor HES-1 prevents mammalian neural differentiation in the central nervous system. EMBO J 13(8):1799–1805PubMedGoogle Scholar
  22. Jesell TM (2000) Neuronal specification in the spinal cord: inductive signals and transcriptional codes. Nat Rev Genet 1:20–29CrossRefGoogle Scholar
  23. Kageyama R, Ohtsuka T, Hatakeyama J, Ohsawa R (2005) Roles of bHLH genes in neural stem cell differentiation. Exp Cell Res 306:343–348CrossRefPubMedGoogle Scholar
  24. Lindsell CE, Boulter J, diSibio G, Gossler A, Weinmaster G (1996) Expression patterns of jagged, Delta1, Notch1, Notch2, and Notch3 genes identify ligand–receptor pairs that may function in neural development. Mol Cell Neurosci 8:14–27CrossRefPubMedGoogle Scholar
  25. Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2∆∆Ct method. Methods 25:402–408CrossRefPubMedGoogle Scholar
  26. Lobe CG (1997) Expression of the helix-loop-helix factor, Hes3, during embryo development suggests a role in early midbrain–hindbrain patterning. Mech Dev 62:227–237CrossRefPubMedGoogle Scholar
  27. Louvi A, Artavanis-Tsakonas S (2006) Notch signalling in vertebrate neural development. Nat Rev Neurosci 7:93–102CrossRefPubMedGoogle Scholar
  28. McDermott KW, Lantos PL (1989) The distribution of glial fibrillary acidic protein and vimentin in postnatal marmoset (Callithrix jacchus) brain. Dev Brain Res 45:169–177CrossRefGoogle Scholar
  29. Morrison SJ, Perez SE, Qiao Z, Verdi JM, Hicks C, Weinmaster G, Anderson DJ (2000) Transient Notch activation initiates an irreversible switch from neurogenesis to gliogenesis by neural crest stem cells. Cell 101:499–510CrossRefPubMedGoogle Scholar
  30. Rath MF, Bailey MJ, Kim JS, Coon SL, Klein DC, Møller M (2009) Developmental and daily expression of the Pax4 and Pax6 homeobox genes in the rat retina: localization of Pax4 in photoreceptor cells. J Neurochem 108:285–294CrossRefPubMedGoogle Scholar
  31. Reaume AG, Conlon RA, Zirngibl R, Yamaguchi TP, Rossant J (1992) Expression analysis of a Notch homologue in the mouse embryo. Dev Biol 154:377–387CrossRefPubMedGoogle Scholar
  32. Scardigli R, Bäumer N, Gruss P, Guillemot F, Le Roux I (2003) Direct and concentration-dependent regulation of the proneural gene Neurogenin2 by Pax6. Development 130:3269–3281CrossRefPubMedGoogle Scholar
  33. Sugimori M, Nagao M, Bertrand N, Parras CM, Guillemot F, Nakafuku M (2007) Combinatorial actions of patterning and HLH transcription factors in the spatiotemporal control of neurogenesis and gliogenesis in the developing spinal cord. Development 134:1617–1629CrossRefPubMedGoogle Scholar
  34. Tanigaki K, Nogaki F, Takahashi J, Tashiro K, Kurooka H, Hono T (2001) Notch1 and Notch3 instructively restrict bFGF-responsive multipotent neural progenitor cells to an astroglial fate. Neuron 29:45–55CrossRefGoogle Scholar
  35. Taylor MK, Yeager K, Morrison SJ (2007) Physiological Notch signalling promotes gliogenesis in the developing peripheral and central nervous systems. Development 134:2435–2447CrossRefPubMedGoogle Scholar
  36. Tokunaga A, Kohyama J, Yoshida T, Nakao K, Sawamoto K, Okano H (2004) Mapping spatio-temporal activation of Notch signaling during neurogenesis and gliogenesis in the developing mouse brain. J Neurochem 90(1):142–154CrossRefPubMedGoogle Scholar
  37. Weinmaster G, Roberts VJ, Lemke G (1991) A homolog of Drosophila Notch expressed during mammalian development. Development 113:199–205PubMedGoogle Scholar
  38. Wu Y, Liu Y, Levine EM, Rao MS (2003) Hes1 but not Hes5 regulates an astrocyte versus oligodendrocyte fate choice in glial restricted precursors. Dev Dynam 226:675–689CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  1. 1.Department of Anatomy, BioSciences InstituteUniversity College CorkCorkIreland

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