Chemogenetic emulation of intraneuronal oxidative stress affects synaptic plasticity

Andrei L Kalinichenko; David Jappy; Georgy M Solius; Dmitry I Maltsev; Yulia A Bogdanova; Liana F Mukhametshina; Rostislav A Sokolov; Aleksandr A Moshchenko; Vladimir A Shaydurov; Andrei V Rozov; Oleg V Podgorny; Vsevolod V Belousov

doi:10.1016/j.redox.2023.102604

Chemogenetic emulation of intraneuronal oxidative stress affects synaptic plasticity

Redox Biol. 2023 Apr:60:102604. doi: 10.1016/j.redox.2023.102604. Epub 2023 Jan 10.

Affiliations

¹ Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, 117997, Moscow, Russia; Lomonosov Moscow State University, 119991, Moscow, Russia.
² Institute of Fundamental Neurology, Federal Center of Brain Research and Neurotechnologies, Federal Medical Biological Agency, 117997, Moscow, Russia; Kazan Federal University, 420008, Kazan, Russia.
³ Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, 117997, Moscow, Russia.
⁴ Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, 117997, Moscow, Russia; Institute of Fundamental Neurology, Federal Center of Brain Research and Neurotechnologies, Federal Medical Biological Agency, 117997, Moscow, Russia.
⁵ Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, 117997, Moscow, Russia; Pirogov Russian National Research Medical University, 117997, Moscow, Russia.
⁶ Pirogov Russian National Research Medical University, 117997, Moscow, Russia; Institute of Biology and Biomedicine, Lobachevsky State University of Nizhny Novgorod, 603022, Nizhny Novgorod, Russia.
⁷ Institute of Fundamental Neurology, Federal Center of Brain Research and Neurotechnologies, Federal Medical Biological Agency, 117997, Moscow, Russia.
⁸ Institute of Fundamental Neurology, Federal Center of Brain Research and Neurotechnologies, Federal Medical Biological Agency, 117997, Moscow, Russia; Institute of Higher Nervous Activity and Neurophysiology, Russian Academy of Sciences, 117485, Moscow, Russia.
⁹ Institute of Fundamental Neurology, Federal Center of Brain Research and Neurotechnologies, Federal Medical Biological Agency, 117997, Moscow, Russia; Institute for Physiology and Pathophysiology, University of Heidelberg, 69120, Heidelberg, Germany. Electronic address: rozov1511@gmail.com.
¹⁰ Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, 117997, Moscow, Russia; Institute of Fundamental Neurology, Federal Center of Brain Research and Neurotechnologies, Federal Medical Biological Agency, 117997, Moscow, Russia; Center for Precision Genome Editing and Genetic Technologies for Biomedicine, Pirogov Russian National Research Medical University, 117997, Moscow, Russia. Electronic address: olegpodgorny@inbox.ru.
¹¹ Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, 117997, Moscow, Russia; Institute of Fundamental Neurology, Federal Center of Brain Research and Neurotechnologies, Federal Medical Biological Agency, 117997, Moscow, Russia; Center for Precision Genome Editing and Genetic Technologies for Biomedicine, Pirogov Russian National Research Medical University, 117997, Moscow, Russia; Institute for Cardiovascular Physiology, Georg August University Göttingen, D-37075, Göttingen, Germany. Electronic address: belousov@fccps.ru.

Abstract

Oxidative stress, a state of disrupted redox signaling, reactive oxygen species (ROS) overproduction, and oxidative cell damage, accompanies numerous brain pathologies, including aging-related dementia and Alzheimer's disease, the most common neurodegenerative disorder of the elderly population. However, a causative role of neuronal oxidative stress in the development of aging-related cognitive decline and neurodegeneration remains elusive because of the lack of approaches for modeling isolated oxidative injury in the brain. Here, we present a chemogenetic approach based on the yeast flavoprotein d-amino acid oxidase (DAAO) for the generation of intraneuronal hydrogen peroxide (H₂O₂). To validate this chemogenetic tool, DAAO and HyPer7, an ultrasensitive genetically encoded H₂O₂ biosensor, were targeted to neurons. Changes in the fluorescence of HyPer7 upon treatment of neurons expressing DAAO with d-norvaline (D-Nva), a DAAO substrate, confirmed chemogenetically induced production of intraneuornal H₂O₂. Then, using the verified chemogenetic tool, we emulated isolated intraneuronal oxidative stress in acute brain slices and, using electrophysiological recordings, revealed that it does not alter basal synaptic transmission and the probability of neurotransmitter release from presynaptic terminals but reduces long-term potentiation (LTP). Moreover, treating neurons expressing DAAO with D-Nva via the patch pipette also decreases LTP. This observation indicates that isolated oxidative stress affects synaptic plasticity at single cell level. Our results broaden the toolset for studying normal redox regulation in the brain and elucidating the role of oxidative stress to the pathogenesis of cognitive aging and the early stages of aging-related neurodegenerative diseases. The proposed approach is useful for identification of early markers of neuronal oxidative stress and may be used in screens of potential antioxidants effective against neuronal oxidative injury.

Keywords: Brain aging; Hydrogen peroxide; Long-term potentiation; Neurodegeneration; Oxidative stress; Synaptic plasticity; d-amino acid oxidase.

MeSH terms

Aged
Antioxidants / pharmacology
Humans
Hydrogen Peroxide* / pharmacology
Neuronal Plasticity / physiology
Oxidative Stress*
Reactive Oxygen Species / pharmacology

Substances

Hydrogen Peroxide
Reactive Oxygen Species
Antioxidants