Presenilins are not required for Aβ42 production in the early secretory pathway

Abstract

Presenilins 1 and 2 (PS1/PS2) have been suggested to be γ-secretases responsible for the proteolytic cleavage of amyloid precursor protein (APP) to form amyloid-β (Aβ), a protein implicated in the development of Alzheimer's disease. Here we examined whether these presenilins are required for the generation of multiple Aβ species by analyzing the production of several forms of secreted and intracellular Aβ in mouse cells lacking PS1, PS2 or both proteins. Although most Aβ species were abolished in PS1/PS2−/− cells, the production of intracellular Aβ42 generated in the endoplasmic reticulum/intermediate compartment was unaffected by the absence of these proteins, either singly or in combination. These results indicate that production of this pool of Aβ occurs independently of PS1/PS2, and therefore, another γ-secretase activity must be responsible for cleavage of APP within the early secretory compartments.

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Figure 1: ELISA quantification of secreted and intracellular Aβ40 and Aβ42 in cortical neurons derived from PS1−/−, PS2−/−, PS+/+, PS1+/? or hPS1 wild-type rescue mice.
Figure 2: Examination of Aβ production in the early secretory pathway.
Figure 3: Pharmacological retention of APP to the early secretory pathway using Brefeldin A or temperature treatments.
Figure 4: Production of Aβ in PS1/PS2−/− or PS+/+ embryonic stem cells.
Figure 5: Aβ can be detected by mass spectrometry of cell lysates from infected PS1/PS2−/− cells.
Figure 6: Modulation of Aβ production with the proteasome inhibitor lactacystin or γ-secretase inhibitors DFK-167 and 2-Napthyl-VF-CHO.

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Acknowledgements

Monoclonal antibodies for the Aβ sandwich ELISA were provided by N. Suzuki and Tekeda Pharmaceuticals, and the calnexin antibody was a gift of A. Helenius. PS1 rescue mice were provided by S. Qian, PS1/PS2−/− and PS+/+ embryonic stem cells by D. Donoviel and B. Yankner, and the APPMV plasmid by C. Liu. We thank R. Stefani and the University of Pennsylvania Protein Chemistry Laboratory as well as J. Souza for mass spectrometry analysis, and B. Giasson, D. Skovronsky and J. Trojanowski for thoughtful input. This work was supported by grants from the National Institute on Aging. C.A.W. was a Howard Hughes Predoctoral Fellow, R.W.D. is supported by a Paul Beeson Faculty Scholar Award and V.M.Y.L. is the John H. Ware III professor of Alzheimer's research.

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Correspondence to Virginia M.-Y. Lee.

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Wilson, C., Doms, R., Zheng, H. et al. Presenilins are not required for Aβ42 production in the early secretory pathway. Nat Neurosci 5, 849–855 (2002). https://doi.org/10.1038/nn898

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