In vivo dynamics of acidosis and oxidative stress in the acute phase of an ischemic stroke in a rodent model

Ilya V Kelmanson; Arina G Shokhina; Daria A Kotova; Matvei S Pochechuev; Alexandra D Ivanova; Alexander I Kostyuk; Anastasiya S Panova; Anastasia A Borodinova; Maxim A Solotenkov; Evgeny A Stepanov; Roman I Raevskii; Aleksandr A Moshchenko; Valeriy V Pak; Yulia G Ermakova; Gijsbert J C van Belle; Viktor Tarabykin; Pavel M Balaban; Ilya V Fedotov; Andrei B Fedotov; Marcus Conrad; Ivan Bogeski; Dörthe M Katschinski; Thorsten R Doeppner; Mathias Bähr; Aleksei M Zheltikov; Vsevolod V Belousov; Dmitry S Bilan

doi:10.1016/j.redox.2021.102178

In vivo dynamics of acidosis and oxidative stress in the acute phase of an ischemic stroke in a rodent model

Redox Biol. 2021 Nov 3:48:102178. doi: 10.1016/j.redox.2021.102178. Online ahead of print.

Authors

Ilya V Kelmanson¹, Arina G Shokhina¹, Daria A Kotova², Matvei S Pochechuev³, Alexandra D Ivanova⁴, Alexander I Kostyuk², Anastasiya S Panova², Anastasia A Borodinova⁵, Maxim A Solotenkov³, Evgeny A Stepanov⁶, Roman I Raevskii², Aleksandr A Moshchenko⁷, Valeriy V Pak², Yulia G Ermakova⁸, Gijsbert J C van Belle⁹, Viktor Tarabykin¹⁰, Pavel M Balaban⁵, Ilya V Fedotov¹¹, Andrei B Fedotov⁶, Marcus Conrad¹², Ivan Bogeski⁹, Dörthe M Katschinski⁹, Thorsten R Doeppner¹³, Mathias Bähr¹⁴, Aleksei M Zheltikov¹¹, Vsevolod V Belousov¹⁵, Dmitry S Bilan¹⁶

Affiliations

¹ M.M. Shemyakin and Yu.A. Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow, 117997, Russia; Laboratory of Experimental Oncology, Pirogov Russian National Research Medical University, 117997, Moscow, Russia.
² M.M. Shemyakin and Yu.A. Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow, 117997, Russia.
³ Physics Department, International Laser Center, M.V. Lomonosov Moscow State University, Moscow, 119992, Russia.
⁴ M.M. Shemyakin and Yu.A. Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow, 117997, Russia; Biological Department, M.V. Lomonosov Moscow State University, Moscow, 119992, Russia.
⁵ Institute of Higher Nervous Activity and Neurophysiology, Russian Academy of Sciences, Moscow, 117485, Russia.
⁶ Physics Department, International Laser Center, M.V. Lomonosov Moscow State University, Moscow, 119992, Russia; Russian Quantum Center, Skolkovo, Moscow Region, 143025, Russia.
⁷ Federal Center of Brain Research and Neurotechnologies, Federal Medical Biological Agency, Moscow, 117997, Russia.
⁸ European Molecular Biology Laboratory, Heidelberg, 69117, Germany.
⁹ Institute for Cardiovascular Physiology, University Medical Center Göttingen, Georg-August-University, Humboldtallee 23, 37073, Göttingen, Germany.
¹⁰ Institute of Cell Biology and Neurobiology, Charité - Universitätsmedizin Berlin, Berlin, 10117, Germany.
¹¹ Physics Department, International Laser Center, M.V. Lomonosov Moscow State University, Moscow, 119992, Russia; Russian Quantum Center, Skolkovo, Moscow Region, 143025, Russia; Kazan Quantum Center, A.N. Tupolev Kazan National Research Technical University, Kazan, 420126, Russia; Department of Physics and Astronomy, Texas A&M University, College Station, TX, 77843, USA.
¹² Laboratory of Experimental Oncology, Pirogov Russian National Research Medical University, 117997, Moscow, Russia; Helmholtz Zentrum München, Institute of Metabolism and Cell Death, Ingolstädter Landstr. 1, Neuherberg, 85764, Germany.
¹³ Department of Neurology, University Medical Center Göttingen, Göttingen, 37075, Germany; Istanbul Medipol University, Research Institute for Health Sciences and Technologies (SABITA), Istanbul, Turkey; Istanbul Medipol University, School of Medicine, Dept. of Physiology, Istanbul, Turkey.
¹⁴ Department of Neurology, University Medical Center Göttingen, Göttingen, 37075, Germany.
¹⁵ M.M. Shemyakin and Yu.A. Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow, 117997, Russia; Laboratory of Experimental Oncology, Pirogov Russian National Research Medical University, 117997, Moscow, Russia; Federal Center of Brain Research and Neurotechnologies, Federal Medical Biological Agency, Moscow, 117997, Russia; Institute for Cardiovascular Physiology, University Medical Center Göttingen, Georg-August-University, Humboldtallee 23, 37073, Göttingen, Germany. Electronic address: belousov@fccps.ru.
¹⁶ M.M. Shemyakin and Yu.A. Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow, 117997, Russia; Laboratory of Experimental Oncology, Pirogov Russian National Research Medical University, 117997, Moscow, Russia. Electronic address: d.s.bilan@gmail.com.

Abstract

Ischemic cerebral stroke is one of the leading causes of death and disability in humans. However, molecular processes underlying the development of this pathology remain poorly understood. There are major gaps in our understanding of metabolic changes that occur in the brain tissue during the early stages of ischemia and reperfusion. In particular, it is generally accepted that both ischemia (I) and reperfusion (R) generate reactive oxygen species (ROS) that cause oxidative stress which is one of the main drivers of the pathology, although ROS generation during I/R was never demonstrated in vivo due to the lack of suitable methods. In the present study, we record for the first time the dynamics of intracellular pH and H₂O₂ during I/R in cultured neurons and during experimental stroke in rats using the latest generation of genetically encoded biosensors SypHer3s and HyPer7. We detect a buildup of powerful acidosis in the brain tissue that overlaps with the ischemic core from the first seconds of pathogenesis. At the same time, no significant H₂O₂ generation was found in the acute phase of ischemia/reperfusion. HyPer7 oxidation in the brain was detected only 24 h later. Comparison of in vivo experiments with studies on cultured neurons under I/R demonstrates that the dynamics of metabolic processes in these models significantly differ, suggesting that a cell culture is a poor predictor of metabolic events in vivo.

Keywords: Genetically encoded fluorescent biosensors; Hydrogen peroxide; Hypoxia/reoxygenation; In vivo optical brain interrogation; Ischemia/reperfusion; Ischemic stroke.