Elsevier

Neuroscience

Volume 153, Issue 3, 15 May 2008, Pages 721-732
Neuroscience

Neuroanatomy
Purkinje cell subtype specification in the cerebellar cortex: Early B-cell factor 2 acts to repress the zebrin II-positive Purkinje cell phenotype

https://doi.org/10.1016/j.neuroscience.2008.01.090Get rights and content

Abstract

The mammalian cerebellar cortex is highly compartmentalized. First, it is subdivided into four transverse expression domains: the anterior zone (AZ), the central zone (CZ), the posterior zone (PZ), and the nodular zone (NZ). Within each zone, the cortex is further subdivided into a symmetrical array of parasagittal stripes. The most extensively studied compartmentation antigen is zebrin II/aldolase c, which is expressed by a subset of Purkinje cells forming parasagittal stripes. Stripe phenotypes are specified early in cerebellar development, in part through the action of early B-cell factor 2 (Ebf2), a member of the atypical helix–loop–helix transcription factor family Collier/Olf1/EBF. In the murine cerebellum, Ebf2 expression is restricted to the zebrin II-immunonegative (zebrin II−) Purkinje cell population. We have identified multiple cerebellar defects in the Ebf2 null mouse involving a combination of selective Purkinje cell death and ectopic expression of multiple genes normally restricted to the zebrin II− subset. The nature of the cerebellar defect in the Ebf2 null is different in each transverse zone. In contrast to the ectopic expression of genes characteristic of the zebrin II+ Purkinje cell phenotype, phospholipase Cβ4 expression, restricted to zebrin II− Purkinje cells in control mice, is well maintained, and the normal number of stripes is present. Taken together, these data suggest that Ebf2 regulates the expression of genes associated with the zebrin II+ Purkinje cell phenotype and that the zebrin II− Purkinje cell subtype is specified independently.

Section snippets

Mice

All animal procedures conformed to institutional regulations and the Guide of the Care and Use of Experimental Animals from the Canadian Council of Animal Care. All experiments conformed to international guidelines on the ethical use of animals. Every effort was made to minimize the number of animals used and their suffering. The targeting construct, described in Corradi et al. (2003), contained a lacZ cDNA and the distribution of lacZ expression during CNS development is in full agreement with

Results

There are two distinct abnormalities in the Ebf2 null cerebellum. First, ectopic gene expression in the zebrin II−/PLCβ4+ population reveals a partial phenotype transdifferentiation from P− to P+. Secondly, the stripe pattern is not normal, and we will argue below that this is likely due to both ectopic gene expression and selective Purkinje cell death (see also Croci et al., 2006). The degree to which each of these effects is observed depends on the particular transverse zone.

Discussion

In this study, we have further characterized the role of Ebf2 in the development of cerebellar topography. The new findings are (i) Ebf2 gene deletion results in the ectopic expression of several characteristic P+ Purkinje cell markers in P− Purkinje cells; (ii) abnormal cerebellar topography in Ebf2 null mice differs between transverse zones: selective Purkinje cell death occurs in the PZ and AZ vermis, but not in the CZ and NZ vermis, and ectopic gene expression occurs in the PZ hemisphere

Acknowledgments

These studies were supported by grants from the Canadian Institutes of Health Research (R.H.), and the CARIPLO foundation (G.G.C.).

References (60)

  • A.H. Ahn et al.

    The cloning of zebrin II reveals its identity with aldolase C

    Development

    (1994)
  • A. Akintunde et al.

    External cuneocerebellar projections and Purkinje cell zebrin II bands: a direct comparison of parasagittal banding in the mouse cerebellum

    J Chem Neuroanat

    (1994)
  • C. Armstrong et al.

    Pattern formation in the cerebellar cortex

    Biochem Cell Biol

    (2000)
  • C. Armstrong et al.

    Selective Purkinje cell ectopia in the cerebellum of the weaver mouse

    J Comp Neurol

    (2001)
  • C. Armstrong et al.

    Constitutive expression of the 25kDa heat shock protein Hsp25 reveals novel parasagittal bands of Purkinje cells in the adult mouse cerebellar cortex

    J Comp Neurol

    (2000)
  • C. Armstrong et al.

    Expression of heat-shock protein Hsp25 in mouse Purkinje cells during development reveals novel features of cerebellar compartmentation

    J Comp Neurol

    (2001)
  • H. Baumeister et al.

    Identification of NFI-binding sites and cloning of NFI-cDNAs suggest a regulatory role for NFI transcription factors in olfactory neuron gene expression

    Brain Res Mol Brain Res

    (1999)
  • G. Brochu et al.

    Zebrin II: a polypeptide antigen expressed selectively by Purkinje cells reveals compartments in rat and fish cerebellum

    J Comp Neurol

    (1990)
  • S.H. Chung et al.

    Compartmentation of GABAB receptor 2 in the mouse cerebellar cortex

    Cerebellum

    (2007)
  • S. Chung et al.

    The anatomy of the cerebellar nuclei in the normal and scrambler mouse as revealed by the expression of the microtubule-associated protein kinesin light chain 3

    Brain Res

    (2007)
  • A. Corradi et al.

    Hypogonadotropic hypogonadism and peripheral neuropathy in Ebf2-null mice

    Development

    (2003)
  • L. Croci et al.

    A key role for the HLH transcription factor EBF2COE2,/E-3 in Purkinje neuron migration and cerebellar cortical topography

    Development

    (2006)
  • M. Crozatier et al.

    Requirement for the Drosophila COE transcription factor Collier in formation of an embryonic muscle: transcriptional response to notch signalling

    Development

    (1999)
  • G. D'Arcangelo et al.

    Reelin is a secreted glycoprotein recognized by the CR-50 monoclonal antibody

    J Neurosci

    (1997)
  • L. Dubois et al.

    XCoe2, a transcription factor of the Col/Olf-1/EBF family involved in the specification of primary neurons in Xenopus

    Curr Biol

    (1998)
  • M.A. Edwards et al.

    Purkinje cell compartments in the reeler mutant mouse as revealed by zebrin II and 9O-acetylated glycolipid antigen expression

    Anat Embryol

    (1994)
  • L.M. Eisenman et al.

    Antigenic compartmentation in the mouse cerebellar cortex: Zebrin and HNK-1 reveal a complex, overlapping molecular topography

    J Comp Neurol

    (1993)
  • J.M. Fritschy et al.

    GABAB-receptor splice variants GB1a and GB1b in rat brain: developmental regulation, cellular distribution and extrasynaptic location

    Eur J Neurosci

    (1999)
  • E. Gallagher et al.

    Cerebellar abnormalities in the disabled (mdab1–1) mouse

    J Comp Neurol

    (1998)
  • S. Garel et al.

    Ebf1 controls early cell differentiation in the embryonic striatum

    Development

    (1999)
  • S. Garel et al.

    Control of the migratory pathway of facial branchiomotor neurones

    Development

    (2000)
  • K. Hashimoto et al.

    Climbing fiber synapse elimination during postnatal cerebellar development requires signal transduction involving G alpha q and phospholipase C beta 4

    Prog Brain Res

    (2000)
  • M. Hashimoto et al.

    Mediolateral compartmentalization of the cerebellum is determined on the “birth date” of Purkinje cells

    J Neurosci

    (2002)
  • R. Hawkes

    An anatomical model of cerebellar modules

    Prog Brain Res

    (1997)
  • R. Hawkes et al.

    Zebrins: molecular markers of compartmentation in the cerebellum

  • R. Hawkes et al.

    Stripes and zones: the origins of regionalization of the adult cerebellum

    Perspect Dev Neurobiol

    (1997)
  • K. Herrup et al.

    The compartmentalization of the cerebellum

    Annu Rev Neurosci

    (1997)
  • B.W. Howell et al.

    Neuronal position in the developing brain is regulated by mouse disabled-1

    Nature

    (1997)
  • Z. Ji et al.

    Topography of Purkinje cell compartments and mossy fiber terminal fields in lobules II and III of the rat cerebellar cortex: spinocerebellar and cuneocerebellar projections

    Neuroscience

    (1994)
  • H. Jiang et al.

    Phospholipase C β4 is involved in modulating the visual response in mice

    Proc Natl Acad Sci U S A

    (1996)
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