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Cytoplasmic and mitochondrial protein translation in axonal and dendritic terminal arborization

Nature Neuroscience volume 10, pages 828–837 (2007)Cite this article

Abstract

We identified a mutation in Aats-gly (also known as gars or glycyl-tRNA synthetase), the Drosophila melanogaster ortholog of the human GARS gene that is associated with Charcot-Marie-Tooth neuropathy type 2D (CMT2D), from a mosaic genetic screen. Loss of gars in Drosophila neurons preferentially affects the elaboration and stability of terminal arborization of axons and dendrites. The human and Drosophila genes each encode both a cytoplasmic and a mitochondrial isoform. Using additional mutants that selectively disrupt cytoplasmic or mitochondrial protein translation, we found that cytoplasmic protein translation is required for terminal arborization of both dendrites and axons during development. In contrast, disruption of mitochondrial protein translation preferentially affects the maintenance of dendritic arborization in adults. We also provide evidence that human GARS shows equivalent functions in Drosophila, and that CMT2D causal mutations show loss-of-function properties. Our study highlights different demands of protein translation for the development and maintenance of axons and dendrites.

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Figure 1: Projection neurons homozygous for team have severely reduced dendritic and axonal terminal arborization.
Figure 2: Mutations in gars cause the team phenotype.
Figure 3: Developmental studies of garsEX34/EX34 phenotypes.
Figure 4: Drosophila GARS localizes to the cytoplasm and mitochondria in projection neurons and Cos-7 cells.
Figure 5: Cytoplasmic protein translation is required for the dendritic and axonal terminal arborization during development.
Figure 6: Projection neuron clones defective for mitochondrial protein translation show progressive defects in dendritic, but not axonal, terminals.
Figure 7: Dendritic and axonal phenotypes in MB γ neurons homozygous for garsEX34, wars4 and tko3.
Figure 8: Function of human GARS in Drosophila projection neurons.

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Acknowledgements

We thank R.J. Watts, E.D. Hoopfer and O. Schuldiner for contributions to mosaic genetic screening; H.T. Jocobs, D.J. Andrew, the Bloomington Drosophila Stock Center and the Kyoto Drosophila Genetic Resource Center for fly stocks; D. Berdnik, T. Komiyama, O. Schuldiner, B. Tasic and H. Zhu for comments on the manuscripts, and M. Miura for supporting T.C. to complete this work. T.C. was a recipient of a Overseas Research Fellowship from Japan Science and Technology Agency and a Postdoctoral Fellowship for Research Abroad from Japan Society for the Promotion of Science. This work was supported by US National Institutes of Health grant R01-DC005982 to L.L. and by the Sumitomo Foundation and a grant from Japan Society for the Promotion of Science to T.C. L.L. is an investigator of the Howard Hughes Medical Institute.

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Authors and Affiliations

  1. Department of Biological Sciences, Howard Hughes Medical Institute, 385 Serra Mall, Stanford University, Stanford, 94305, California, USA

    Takahiro Chihara, David Luginbuhl & Liqun Luo

  2. Department of Genetics, Graduate School of Pharmaceutical Sciences, University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, 113-0033, Tokyo, Japan

    Takahiro Chihara

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Contributions

T.C. designed the study with the help of L.L. T.C. conducted the experimental work and analyzed the data, D.L. assisted with the forward genetic screen, and T.C. and L.L. wrote the manuscript.

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Correspondence to Liqun Luo.

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Supplementary information

Supplementary Fig. 1

Lack of axon terminal arborization in team−/− projection neurons. (PDF 794 kb)

Supplementary Fig. 2

MARCM rescue experiments of garsEX34/EX34 projection neuron clones with either cytoplasmic or mitochondrial GARS. (PDF 3795 kb)

Supplementary Fig. 3

Generation of Df(3L)mito. (PDF 120 kb)

Supplementary Methods (PDF 102 kb)

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Chihara, T., Luginbuhl, D. & Luo, L. Cytoplasmic and mitochondrial protein translation in axonal and dendritic terminal arborization. Nat Neurosci 10, 828–837 (2007). https://doi.org/10.1038/nn1910

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