Kinetic monitoring of neuronal stress response to proteostasis dysfunction

• Authors: Angel J Santiago-Lopez, Emory University; Ken Berglund, Emory University; Robert Gross, Emory University; Claire-Anne Gutekunst, Emory University
• Journal Title: MOLECULAR AND CELLULAR NEUROSCIENCE
• Volume: Volume 118
• Publisher: ACADEMIC PRESS INC ELSEVIER SCIENCE | 2021-11-19, Pages 103682-103682
• Type of Work: Article | Post-print: After Peer Review
• Publisher DOI: http://doi.org/10.1016/j.mcn.2021.103682
• Subjects: Health Sciences, Medicine and Surgery; Engineering, Biomedical
• Keywords: Science & Technology; Life Sciences & Biomedicine; Neurosciences; Neurosciences & Neurology; Neurodegeneration; Integrated stress response; Proteostasis; Unfolded protein response; Alpha-synuclein; UNFOLDED PROTEIN RESPONSE; ALPHA-SYNUCLEIN; PARKINSONS-DISEASE; GENE-THERAPY; LEWY BODIES; ER STRESS; IN-VITRO; PROTEASOME; IMPAIRMENT; MODEL

Abstract

Proteostasis dysfunction and activation of the unfolded protein response (UPR) are characteristic of all major neurodegenerative diseases. Nevertheless, although the UPR and proteostasis dysfunction has been studied in great detail in model organisms like yeast and mammalian cell lines, it has not yet been examined in neurons. In this study, we applied a viral vector-mediated expression of a reporter protein based on a UPR transcription factor, ATF4, and time-lapse fluorescent microscopy to elucidate how mouse primary neurons respond to pharmacological and genetic perturbations to neuronal proteostasis. In in vitro models of endoplasmic reticulum (ER) stress and proteasome inhibition, we used the ATF4 reporter to reveal the time course of the neuronal stress response relative to neurite degeneration and asynchronous cell death. We showed how potential neurodegenerative disease co-factors, ER stress and mutant α-synuclein overexpression, impacted neuronal stress response and overall cellular health. This work therefore introduces a viral vector-based reporter that yields a quantifiable readout suitable for non-cell destructive kinetic monitoring of proteostasis dysfunction in neurons by harnessing ATF4 signaling as part of the UPR activation.