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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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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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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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)
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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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DOI: https://doi.org/10.1038/nn1910
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