Abstract
Down syndrome (DS) is characterized by the trisomy of chromosome 21 and by cognitive deficits that have been related to neuronal morphological alterations in humans, as well as in animal models. The gene encoding for amyloid precursor protein (APP) is present in autosome 21, and its overexpression in DS has been linked to neuronal dysfunction, cognitive deficit, and Alzheimer’s disease-like dementia. In particular, the neuronal ability to extend processes and branching is affected. Current evidence suggests that APP could also regulate neurite growth through its role in the actin cytoskeleton, in part by influencing p21-activated kinase (PAK) activity. The latter effect is carried out by an increased abundance of the caspase cleavage-released carboxy-terminal C31 fragment. In this work, using a neuronal cell line named CTb, which derived from the cerebral cortex of a trisomy 16 mouse, an animal model of human DS, we observed an overexpression of APP, elevated caspase activity, augmented cleavage of the C-terminal fragment of APP, and increased PAK1 phosphorylation. Morphometric analyses showed that inhibition of PAK1 activity with FRAX486 increased the average length of the neurites, the number of crossings per Sholl ring, the formation of new processes, and stimulated the loss of processes. Considering our results, we propose that PAK hyperphosphorylation impairs neurite outgrowth and remodeling in the cellular model of DS, and therefore we suggest that PAK1 may be a potential pharmacological target.
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This work was funded by FONDECYT Grant #1130241, 1161450 (Chile) to PC, the Fondation pour la Recherche sur le Cerveau (FRC) and the Fondation Jérôme Lejeune (J-V. B., France), CONICYT for funding of Basal Centre, CeBiB, FB0001 and P09-022-F from ICM-ECONOMIA (Chile), and CONICYT grant #21120728 (Chile).
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N.B. carried out most experiments and wrote the main manuscript. R. P-N. assisted with blotting experiments and data analysis. B.G. conducted part of FRAX486 inhibition experiments. A. B. designed the algorithms for morphological analysis. C.A. assisted with blotting experiments and morphological analysis. K.P. and V.J. assisted with blotting experiments related to APP Δ665 detection and caspase activity. J-V. B. assisted in PAK1 phosphorylation and activity experiments. A.M. C assisted in planning, data analysis, and manuscript text. P.C. assisted in all aforementioned aspects of the research and writing of the text. All authors reviewed the manuscript and are aware it is being submitted to this journal in its present version.
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Barraza-Núñez, N., Pérez-Núñez, R., Gaete-Ramírez, B. et al. Pharmacological Inhibition of p-21 Activated Kinase (PAK) Restores Impaired Neurite Outgrowth and Remodeling in a Cellular Model of Down Syndrome. Neurotox Res 41, 256–269 (2023). https://doi.org/10.1007/s12640-023-00638-3
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DOI: https://doi.org/10.1007/s12640-023-00638-3