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The Influence of β-Arrestin-2 Gene Knockout in Mice on Survival of Cultured Astrocytes Exposed to Thrombin and on the Cerebral Thrombosis Aftereffects In Vivo

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Biochemistry (Moscow), Supplement Series A: Membrane and Cell Biology Aims and scope

Abstract—

Thrombin is a multifunctional serine protease that attracts the attention of many researchers in particular in connection with a wide range of its effects in the nervous tissue. It is known that the main thrombin receptor is a protease-activated type 1 receptor, the functional activity of which is associated with adapter protein β-arrestin-2. Here we assessed potential involvement of β-arrestin-2 in the thrombin toxicity both in vitro and in vivo using gene knockout mice. It was found that thrombin induced dose-dependent cell death of cultured β-arrestin-2–/– astrocytes 48 h after the exposure. In contrast, thrombin did not exert any effects on the survival of astrocytes from wild-type animals. The in vivo study showed that β-arrestin-2 gene knockout did not alter the severity of the focal photoinduced cerebral ischemia aftereffects, which could involve additional cell types and molecular mechanisms in nervous tissue damage. Our findings demonstrate for the first time the necessity of β-arrestin-2 for the survival of mouse astrocytes under the toxic action of thrombin. At the CNS level, however, further studies are required to determine the key targets of this protease in each cell types of the nervous tissue and to clarify the role of β-arrestin signaling in neuroprotection.

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ACKNOWLEDGMENTS

The authors thank D.A. Gorbachev (Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences) and V.S. Vyushkov (Moscow Lomonosov State University) for the help in mouse genotyping and interpretation of the obtained data. The work was supported by the Russian Foundation for Basic Research (project no. 18-34-00977).

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Authors and Affiliations

  1. Moscow Lomonosov State University, 119234, Moscow, Russia

    M. D. Galkov, M. V. Gulyaev & L. R. Gorbacheva

  2. Pirogov Russian National Research Medical University, 117997, Moscow, Russia

    M. D. Galkov, A. E. Ivanova, I. G. Savinkova & L. R. Gorbacheva

  3. Koltzov Institute of Developmental Biology, Russian Academy of Sciences, 119334, Moscow, Russia

    E. V. Kiseleva

Authors
  1. M. D. Galkov
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  2. A. E. Ivanova
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  3. M. V. Gulyaev
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  4. E. V. Kiseleva
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  5. I. G. Savinkova
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  6. L. R. Gorbacheva
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Correspondence to M. D. Galkov.

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The authors declare that they have no conflict of interest.

All procedures were performed in accordance with Directive 2010/63/EU of the European Parliament and the Council of the European Union. Experimental protocols were approved by the Bioethics Committee of Moscow Lomonosov State University (protocol no. 2018-10-25-93-0-3).

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Translated by M. Galkov

Abbreviations: APC, activated protein C; BBB, blood–brain barrier; GPCRs, G-protein-coupled receptors; GRKs, kinases of G-protein-coupled receptors; MRI, magnetic resonance imaging; PARs, protease-activated receptors; PC, protein C; PCR, polymerase chain reaction; TTC, 2,3,5-triphenyltetrazolium chloride.

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Galkov, M.D., Ivanova, A.E., Gulyaev, M.V. et al. The Influence of β-Arrestin-2 Gene Knockout in Mice on Survival of Cultured Astrocytes Exposed to Thrombin and on the Cerebral Thrombosis Aftereffects In Vivo. Biochem. Moscow Suppl. Ser. A 14, 17–23 (2020). https://doi.org/10.1134/S1990747819060060

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