The Cerebellum

, Volume 7, Issue 3, pp 451–466

Both Cell-Autonomous and Cell Non-Autonomous Functions of GAP-43 are Required for Normal Patterning of the Cerebellum In Vivo

  • Yiping Shen
  • Rashmi Mishra
  • Shyamala Mani
  • Karina F. Meiri
Article

Abstract

Growth-associated protein 43 (GAP-43) is required for development of a functional cerebral cortex in vertebrates; however, its role in cerebellar development is not well understood. Recently, we showed that absence of GAP-43 caused defects in proliferation, differentiation, and polarization of cerebellar granule cells. In this paper, we show that absence of GAP-43 causes defects in cerebellar patterning that reflect both cell-autonomous and non-autonomous functions. Cell-autonomous effects of GAP-43 impact precursor proliferation and axon targeting: In its absence, (1) proliferation of granule cell precursors in response to sonic hedgehog and fibroblast growth factor is inhibited, (2) proliferation of neuroepithelial precursors is inhibited, and (3) targeting of climbing fibers to the central lobe is disrupted. Cell non-autonomous effects of GAP-43 impact differentiated Purkinje cells in which GAP-43 has been downregulated: In its absence, both maturation and mediolateral patterning of Purkinje cells are inhibited. Both cell-autonomous and non-autonomous functions of GAP-43 involve its phosphorylation by protein kinase C. GAP-43 is phosphorylated in granule cell precursors in response to sonic hedgehog in vitro, and phosphorylated GAP-43 is also found in proliferating neuroepithelium and climbing fibers. Phosphorylated GAP-43 is specifically enriched in the presynaptic terminals of parallel and climbing fibers that innervate Purkinje cell bodies and dendrites. The cell-autonomous and non-autonomous effects of GAP-43 converge on the central lobe. The multiple effects of GAP-43 on cerebellar development suggest that it is a critical downstream transducer of signaling mechanisms that integrate generation of cerebellar structure with functional parcellation at the central lobe.

Keywords

GAP-43 Knockout mouse Cerebellar granule cell proliferation Purkinje cell patterning Sonic hedgehog Climbing fibers 

Abbreviations

ABT

anterobasal tract

ADT

anterodorsal tract

AL

anterior lobe

BrDU

bromodeoxyuridine

CeL

central lobe

CeT

central tract

CF

climbing fiber

Cp

cerebellar peduncle

EGL

external granule cell layer

FGF

fibroblast growth factor

FN

fastigial nucleus

GAP-43

growth-associated protein 43

GC

granule cell

GCP

granule cell precursor

IgSFCAM

immunoglobulin superfamily cell adhesion molecule

LI

labeling index

ML

molecular layer

NCAM

neural cell adhesion molecule

PC

Purkinje cell

PCP

Purkinje cell precursor

PKC

protein kinase C

PL

posterior lobe

Ptc-1

patched-1

POT

posterior tract

Shh

sonic hedgehog

Smo

smoothened

SVZ

subventricular zone

References

  1. 1.
    Altman J, Bayer SA (1987) Development of the precerebellar nuclei in the rat: 11. The intramural olivary migratory stream and the neurogenetic organization of the inferior olive. J Comp Neurol 257:490–512PubMedCrossRefGoogle Scholar
  2. 2.
    Altman J, Bayer SA (1996) Development of the cerebellar system. CRC, New YorkGoogle Scholar
  3. 3.
    Armstrong CL, Krueger-Naug AM, Currie RW, Hawkes R (2000) Constitutive expression of the 25-kDa heat shock protein Hsp25 reveals novel parasagittal bands of Purkinje cells in the adult mouse cerebellar cortex. J Comp Neurol 416:383–397PubMedCrossRefGoogle Scholar
  4. 4.
    Blatt GJ, Eisenman LM (1993) The olivocerebellar projection in normal (+/+), heterozygous weaver (wv/+), and homozygous weaver (wv/wv) mutant mice: comparison of terminal pattern and topographic organization. Exp Brain Res 95:187–201PubMedCrossRefGoogle Scholar
  5. 5.
    Borghesani PR, Peyrin JM, Klein R, Rubin J, Carter AR, Schwartz PM, Luster A, Corfas G, Segal RA (2002) BDNF stimulates migration of cerebellar granule cells. Development 129:1435–1442PubMedGoogle Scholar
  6. 6.
    Cavallaro U, Niedermeyer J, Fuxa M, Christofori G (2001) N-CAM modulates tumour-cell adhesion to matrix by inducing FGF-receptor signalling. Nat Cell Biol 3:650–657PubMedCrossRefGoogle Scholar
  7. 7.
    Chan SY, Murakami K, Routtenberg A (1986) Phosphoprotein F1: purification and characterization of a brain kinase C substrate related to plasticity. J Neurosci 6:3618–3627PubMedGoogle Scholar
  8. 8.
    Chedotal A, Pourquie O, Ezan F, San Clemente H, Sotelo C (1996) BEN as a presumptive target recognition molecule during the development of the olivocerebellar system. J Neurosci 16:3296–3310PubMedGoogle Scholar
  9. 9.
    Console-Bram LM, Fitzpatrick-McElligott SG, McElligott JG (1996) Distribution of GAP-43 mRNA in the immature and adult cerebellum: a role for GAP-43 in cerebellar development and neuroplasticity. Brain Res Dev Brain Res 95:97–106PubMedCrossRefGoogle Scholar
  10. 10.
    Corrales JD, Blaess S, Mahoney EM, Joyner AL (2006) The level of sonic hedgehog signaling regulates the complexity of cerebellar foliation. Development 133:1811–1821PubMedCrossRefGoogle Scholar
  11. 11.
    Dahmane N, Ruiz i Altaba A (1999) Sonic hedgehog regulates the growth and patterning of the cerebellum. Development 126:3089–3100PubMedGoogle Scholar
  12. 12.
    DeBernardo AP, Chang S (1996) Heterophilic interactions of DM-GRASP: GRASP-NgCAM interactions involved in neurite extension. J Cell Biol 133:657–666PubMedCrossRefGoogle Scholar
  13. 13.
    Drake-Baumann R (2005) Rapid modulation of inhibitory synaptic currents in cerebellar Purkinje cells by BDNF. Synapse 57:183–190PubMedCrossRefGoogle Scholar
  14. 14.
    Fogarty MP, Emmenegger BA, Grasfeder LL, Oliver TG, Wechsler-Reya RJ (2007) Fibroblast growth factor blocks sonic hedgehog signaling in neuronal precursors and tumor cells. Proc Natl Acad Sci U S A 104:2973–2978PubMedCrossRefGoogle Scholar
  15. 15.
    Gao WO, Heintz N, Hatten ME (1991) Cerebellar granule cell neurogenesis is regulated by cell-cell interactions in vitro. Neuron 6:705–715PubMedCrossRefGoogle Scholar
  16. 16.
    Gerendasy D (1999) Homeostatic tuning of Ca2+ signal transduction by members of the calpacitin protein family. J Neurosci Res 58:107–119PubMedCrossRefGoogle Scholar
  17. 17.
    Goldowitz D, Hamre K (1998) The cells and molecules that make a cerebellum. Trends Neurosci 21:375–382PubMedCrossRefGoogle Scholar
  18. 18.
    Gonzalez-Quevedo R, Shoffer M, Horng L, Oro AE (2005) Receptor tyrosine phosphatase-dependent cytoskeletal remodeling by the hedgehog-responsive gene MIM/BEG4. J Cell Biol 168:453–463PubMedCrossRefGoogle Scholar
  19. 19.
    Guerrero I, Chiang C (2007) A conserved mechanism of hedgehog gradient formation by lipid modifications. Trends Cell Biol 17:1–5PubMedCrossRefGoogle Scholar
  20. 20.
    Hatten ME, Heintz N (1995) Mechanisms of neural patterning and specification in the developing cerebellum. Annu Rev Neurosci 18:385–408PubMedGoogle Scholar
  21. 21.
    He Q, Dent EW, Meiri KF (1997) Modulation of actin filament behavior by GAP-43 (neuromodulin) is dependent on the phosphorylation status of serine 41, the protein kinase C site. J Neurosci 17:3515–3524PubMedGoogle Scholar
  22. 22.
    Herrup K, Kuemerle B (1997) The compartmentalization of the cerebellum. Annu Rev Neurosci 20:61–90PubMedCrossRefGoogle Scholar
  23. 23.
    Heo JS, Lee MY, Han HJ (2007) Sonic Hedgehog stimulates mouse embryonic stem cell proliferation by cooperation of Ca2+/protein kinase C and EGF receptor as well as Gli1 activation. Stem Cells 12:3069–3080CrossRefGoogle Scholar
  24. 24.
    Inouye M, Murakami U (1980) Temporal and spatial patterns of Purkinje cell formation in the mouse cerebellum. J Comp Neurol 194:499–503PubMedCrossRefGoogle Scholar
  25. 25.
    Ito M (2002) The molecular organization of cerebellar long-term depression. Nat Rev Neurosci 3:896–902PubMedCrossRefGoogle Scholar
  26. 26.
    Ito M (2006) Cerebellar circuitry as a neuronal machine. Prog Neurobiol 78:272–303PubMedCrossRefGoogle Scholar
  27. 27.
    Jorgensen OS (1994) Neural cell adhesion molecule and D3 protein in the cerebellum of weaver mutant mice. Int J Dev Neurosci 12:213–225PubMedCrossRefGoogle Scholar
  28. 28.
    Korshunova I, Novitskaya V, Kiryushko D, Pedersen N, Kolkova K, Kropotova E, Mosevitsky M, Rayko M, Morrow JS, Ginzburg I, Berezin V, Bock E (2007) GAP-43 regulates NCAM-180-mediated neurite outgrowth. J Neurochem 100:1599–1612PubMedGoogle Scholar
  29. 29.
    Koyner J, Demarest K, McCaughran J Jr, Cipp L, Hitzemann R (2000) Identification and time dependence of quantitative trait loci for basal locomotor activity in the BXD recombinant inbred series and a B6D2 F2 intercross. Behav Genet 3:159–170CrossRefGoogle Scholar
  30. 30.
    Larsson C (2006) Protein kinase C and the regulation of the actin cytoskeleton. Cell Signal 18:276–284PubMedCrossRefGoogle Scholar
  31. 31.
    Larouche M, Hawkes R (2006) From clusters to stripes: the developmental origins of adult cerebellar compartmentation. Cerebellum 5:77–88PubMedCrossRefGoogle Scholar
  32. 32.
    Larouche M, Che PM, Hawkes R (2006) Neurogranin expression identifies a novel array of Purkinje cell parasagittal stripes during mouse cerebellar development. J Comp Neurol 494:215–227PubMedCrossRefGoogle Scholar
  33. 33.
    Laux T, Fukami K, Thelen M, Golub T, Frey D, Caroni P (2000) GAP43, MARCKS, and CAP23 modulate PI(4,5)P(2) at plasmalemmal rafts, and regulate cell cortex actin dynamics through a common mechanism. J Cell Biol 149:1455–1472PubMedCrossRefGoogle Scholar
  34. 34.
    Leshchyns’ka I, Sytnyk V, Morrow JS, Schachner M (2003) Neural cell adhesion molecule (NCAM) association with PKCbeta2 via betaI spectrin is implicated in NCAM-mediated neurite outgrowth. J Cell Biol 161:625–639PubMedCrossRefGoogle Scholar
  35. 35.
    Lustig RH, Hua P, Wilson MC, Federoff HJ (1993) Ontogeny, sex dimorphism, and neonatal sex hormone determination of synapse-associated messenger RNAs in rat brain. Brain Res Mol Brain Res 20:101–110PubMedCrossRefGoogle Scholar
  36. 36.
    Machold R, Fishell G (2005) Math1 is expressed in temporally discrete pools of cerebellar rhombic-lip neural progenitors. Neuron 48:17–24PubMedCrossRefGoogle Scholar
  37. 37.
    Maier DL, Mani S, Donovan SL, Soppet D, Tessarollo L, McCasland JS, Meiri KF (1999) Disrupted cortical map and absence of cortical barrels in growth-associated protein (GAP)-43 knockout mice. Proc Natl Acad Sci U S A 96:9397–9402PubMedCrossRefGoogle Scholar
  38. 38.
    Mani S, Shen Y, Schaefer J, Meiri KF (2001) Failure to express GAP-43 during neurogenesis affects cell cycle regulation and differentiation of neural precursors and stimulates apoptosis of neurons. Mol Cell Neurosci 17:54–66PubMedCrossRefGoogle Scholar
  39. 39.
    Maricich SM, Herrup K (1999) Pax-2 expression defines a subset of GABAergic interneurons and their precursors in the developing murine cerebellum. J Neurobiol 41:281–294PubMedCrossRefGoogle Scholar
  40. 40.
    Meiri KF, Bickerstaff LE, Schwob JE (1991) Monoclonal antibodies show that kinase C phosphorylation of GAP-43 during axonogenesis is both spatially and temporally restricted in vivo. J Cell Biol 112:991–1005PubMedCrossRefGoogle Scholar
  41. 41.
    Meiri KF, Saffell JL, Walsh FS, Doherty P (1998) Neurite outgrowth stimulated by neural cell adhesion molecules requires growth-associated protein-43 (GAP-43) function and is associated with GAP-43 phosphorylation in growth cones. J Neurosci 18:10429–10437PubMedGoogle Scholar
  42. 42.
    Metz GA, Schwab ME (2004) Behavioral characterization in a comprehensive mouse test battery reveals motor and sensory impairments in growth-associated protein-43 null mutant mice. Neuroscience 129:563–574PubMedCrossRefGoogle Scholar
  43. 43.
    Mishra R, Gupta SK, Meiri KF, Fong M, Thostrup P, Juncker D, Mani S (2008) GAP-43 is key to mitotic spindle control and centrosome-based polarization in neurons. Cell Cycle 7:348–357PubMedGoogle Scholar
  44. 44.
    Molinari HH, Schultze KE, Strominger NL (1996) Gracile, cuneate, and spinal trigeminal projections to inferior olive in rat and monkey. J Comp Neurol 375:467–480PubMedCrossRefGoogle Scholar
  45. 45.
    Niethammer P, Delling M, Sytnyk V, Dityatev A, Fukami K, Schachner M (2002) Cosignaling of NCAM via lipid rafts and the FGF receptor is required for neuritogenesis. J Cell Biol 157:521–532PubMedCrossRefGoogle Scholar
  46. 46.
    Ozol K, Hayden JM, Oberdick J, Hawkes R (1999) Transverse zones in the vermis of the mouse cerebellum. J Comp Neurol 412:95–111PubMedCrossRefGoogle Scholar
  47. 47.
    Pijpers A, Apps R, Pardoe J, Voogd J, Ruigrok TJ (2006) Precise spatial relationships between mossy fibers and climbing fibers in rat cerebellar cortical zones. J Neurosci 26:12067–12080PubMedCrossRefGoogle Scholar
  48. 48.
    Rekart JL, Meiri K, Routtenberg A (2005) Hippocampal-dependent memory is impaired in heterozygous GAP-43 knockout mice. Hippocampus 15:1–7PubMedCrossRefGoogle Scholar
  49. 49.
    Routtenberg A, Cantallops I, Zaffuto S, Serrano P, Namgung U (2000) Enhanced learning after genetic overexpression of a brain growth protein. Proc Natl Acad Sci U S A 97:7657–7662PubMedCrossRefGoogle Scholar
  50. 50.
    Rubin JB, Choi Y, Segal RA (2002) Cerebellar proteoglycans regulate sonic hedgehog responses during development. Development 129:2223–2232PubMedGoogle Scholar
  51. 51.
    Schwartz PM, Borghesani PR, Levy RL, Pomeroy SL, Segal RA (1997) Abnormal cerebellar development and foliation in BDNF −/− mice reveals a role for neurotrophins in CNS patterning. Neuron 19:269–281PubMedCrossRefGoogle Scholar
  52. 52.
    Sharma SK, Carew TJ (2002) Inclusion of phosphatase inhibitors during Western blotting enhances signal detection with phospho-specific antibodies. Anal Biochem 307:187–189PubMedCrossRefGoogle Scholar
  53. 53.
    Shen Y, Mani S, Donovan SL, Schwob JE, Meiri KF (2002) GAP-43 is required for commissural axon guidance in the developing vertebrate nervous system. J Neurosci 22:239–247PubMedGoogle Scholar
  54. 54.
    Shughrue PJ, Dorsa DM (1994) The ontogeny of GAP-43 (neuromodulin) mRNA in postnatal rat brain: evidence for a sex dimorphism. J Comp Neurol 340:174–184PubMedCrossRefGoogle Scholar
  55. 55.
    Sotelo C (2004) Cellular and genetic regulation of the development of the cerebellar system. Prog Neurobiol 72:295–339PubMedCrossRefGoogle Scholar
  56. 56.
    Stricker SH, Meiri K, Götz M (2006) P-GAP-43 is enriched in horizontal cell divisions throughout rat cortical development. Cereb Cortex 16(Suppl 1):121–131CrossRefGoogle Scholar
  57. 57.
    Sugihara I, Shinoda Y (2004) Molecular, topographic, and functional organization of the cerebellar cortex: a study with combined aldolase C and olivocerebellar labeling. J Neurosci 24:8771–8785PubMedCrossRefGoogle Scholar
  58. 58.
    Taber-Pierce E (1975) Histogenesis of the deep cerebellar nuclei in the mouse: an autoradiographic study. Brain Res 95:503–518CrossRefGoogle Scholar
  59. 59.
    Traiffort E, Charytoniuk D, Watroba L, Faure H, Sales N, Ruat M (1999) Discrete localizations of hedgehog signalling components in the developing and adult rat nervous system. Eur J Neurosci 11:3199–3214PubMedCrossRefGoogle Scholar
  60. 60.
    Wahlsten D, Andison M (1991) Patterns of cerebellar foliation in recombinant inbred mice. Brain Res 557:184–189PubMedCrossRefGoogle Scholar
  61. 61.
    Wallace VA (1999) Purkinje-cell-derived sonic hedgehog regulates granule neuron precursor cell proliferation in the developing mouse cerebellum. Curr Biol 9:445–448PubMedCrossRefGoogle Scholar
  62. 62.
    Wang VY, Rose MF, Zoghbi HY (2005) Math1 expression redefines the rhombic lip derivatives and reveals novel lineages within the brainstem and cerebellum. Neuron 48:31–43PubMedCrossRefGoogle Scholar
  63. 63.
    Wechsler-Reya RJ, Scott MP (1999) Control of neuronal precursor proliferation in the cerebellum by sonic hedgehog. Neuron 22:103–114PubMedCrossRefGoogle Scholar
  64. 64.
    Zhang GR, Wang X, Kong L, Lu XG, Lee B, Liu M, Sun M, Franklin C, Cook RG, Geller AI (2005) Genetic enhancement of visual learning by activation of protein kinase C pathways in small groups of rat cortical neurons. J Neurosci 25:8468–8481PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2008

Authors and Affiliations

  • Yiping Shen
    • 1
  • Rashmi Mishra
    • 2
  • Shyamala Mani
    • 2
  • Karina F. Meiri
    • 1
  1. 1.Department of Anatomy and Cellular BiologyTufts University School of MedicineBostonUSA
  2. 2.National Brain Research CenterManesarIndia

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