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

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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.

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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

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Acknowledgments

We thank Dr William Brunken for initially alerting us to the cerebellar phenotype, Dr Richard Hawkes for helpful discussions and Lilly Nguyen for expert technical assistance. This work was supported by NS33118 (KFM) and an NIH R03TW0605 Fogarty International Award (KFM and SM).

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Authors and Affiliations

  1. Department of Anatomy and Cellular Biology, Tufts University School of Medicine, 136 Harrison Avenue, Boston, MA, 02111, USA

    Yiping Shen & Karina F. Meiri

  2. National Brain Research Center, Manesar, Haryana, 122-050, India

    Rashmi Mishra & Shyamala Mani

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  1. Yiping Shen
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  2. Rashmi Mishra
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  3. Shyamala Mani
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  4. Karina F. Meiri
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Correspondence to Karina F. Meiri.

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Yiping Shen and Rashmi Mishra contributed equally to this work.

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Shen, Y., Mishra, R., Mani, S. et al. Both Cell-Autonomous and Cell Non-Autonomous Functions of GAP-43 are Required for Normal Patterning of the Cerebellum In Vivo. Cerebellum 7, 451–466 (2008). https://doi.org/10.1007/s12311-008-0049-5

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