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APP binds DR6 to trigger axon pruning and neuron death via distinct caspases

Abstract

Naturally occurring axonal pruning and neuronal cell death help to sculpt neuronal connections during development, but their mechanistic basis remains poorly understood. Here we report that β-amyloid precursor protein (APP) and death receptor 6 (DR6, also known as TNFRSF21) activate a widespread caspase-dependent self-destruction program. DR6 is broadly expressed by developing neurons, and is required for normal cell body death and axonal pruning both in vivo and after trophic-factor deprivation in vitro. Unlike neuronal cell body apoptosis, which requires caspase 3, we show that axonal degeneration requires caspase 6, which is activated in a punctate pattern that parallels the pattern of axonal fragmentation. DR6 is activated locally by an inactive surface ligand(s) that is released in an active form after trophic-factor deprivation, and we identify APP as a DR6 ligand. Trophic-factor deprivation triggers the shedding of surface APP in a β-secretase (BACE)-dependent manner. Loss- and gain-of-function studies support a model in which a cleaved amino-terminal fragment of APP (N-APP) binds DR6 and triggers degeneration. Genetic support is provided by a common neuromuscular junction phenotype in mutant mice. Our results indicate that APP and DR6 are components of a neuronal self-destruction pathway, and suggest that an extracellular fragment of APP, acting via DR6 and caspase 6, contributes to Alzheimer’s disease.

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Figure 1: DR6 regulates degeneration of several neuronal classes.
Figure 2: DR6 regulates axon pruning in vitro and in vivo.
Figure 3: BAX and caspase 6 regulate axonal degeneration.
Figure 4: The N terminus of APP is a regulated DR6 ligand.
Figure 5: The APP N terminus regulates degeneration.
Figure 6: APP and DR6 signalling: in vivo evidence, and model.

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Acknowledgements

We thank R. Axel, C. Bargmann, B. de Strooper, V. Dixit, C. Henderson, J. Lewcock, R. Scheller, R. Vassar, R. Watts, and members of the M.T.-L. laboratory for helpful discussions and suggestions, and critical reading of the manuscript, and A. Bruce for making the diagrams. We thank P. Hass and members of his laboratory (Genentech) for generation and purification of the DR6 ectodomain and APP(1–286), and W.-C. Liang and Y. Wu (Genentech) for binding experiments with purified proteins. Supported by Genentech (A.N. and M.T.-L.) and National Eye Institute grant R01 EY07025 (D.D.M.O.’L.).

Author Contributions A.N. performed most of the experiments, with the exception of the analysis of retinal projections and the experiments listed in the Acknowledgements, and co-wrote the paper. The retinotectal analysis was performed by T.M. and supervised by D.D.M.O.’L. M.T.-L. supervised or co-supervised all experiments, and co-wrote the paper.

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Correspondence to Marc Tessier-Lavigne.

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A.N. and M.T.-L. are employees of Genentech, Inc., which has a commercial interest in some of the molecules studied here.

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Nikolaev, A., McLaughlin, T., O’Leary, D. et al. APP binds DR6 to trigger axon pruning and neuron death via distinct caspases. Nature 457, 981–989 (2009). https://doi.org/10.1038/nature07767

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