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Inhibition of Intracellular Type 10 Adenylyl Cyclase Protects Cortical Neurons Against Reperfusion-Induced Mitochondrial Injury and Apoptosis

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Abstract

Mitochondrial injury significantly contributes to the neuronal death under cerebral ischemia and reperfusion. Within several signaling pathways, cyclic adenosine monophosphate (cAMP) signaling plays a substantial role in mitochondrial injury and cell death. Traditionally, the source of cellular cAMP has been attributed to the membrane-bound adenylyl cyclase, whereas the role of the intracellular localized type 10 soluble adenylyl cyclase (sAC) in neuronal pathology has not been considered. Since neurons express an active form of sAC, we aimed to investigate the role of sAC in reperfusion-induced neuronal apoptosis. For this purpose, the in vitro model of oxygen/glucose deprivation (simulated ischemia, 1 h), followed by recovery (simulated reperfusion, 12 h) in rat embryonic neurons, was applied. Although ischemia alone had no significant effect on apoptosis, reperfusion led to an activation of the mitochondrial pathway of apoptosis, hallmarked by mitochondrial depolarization, cytochrome c release, and mitochondrial ROS formation. These effects were accompanied by significantly augmented sAC expression and increased cellular cAMP content during reperfusion. Pharmacological suppression of sAC during reperfusion reduced cellular cAMP and ameliorated reperfusion-induced mitochondrial apoptosis and ROS formation. Similarly, sAC knockdown prevented neuronal death. Further analysis revealed a role of protein kinase A (PKA), a major downstream target of sAC, in reperfusion-induced neuronal apoptosis and ROS formation. In conclusion, the results show a causal role of intracellular, sAC-dependent cAMP signaling in reperfusion-induced mitochondrial injury and apoptosis in neurons. The protective effect of sAC inhibition during the reperfusion phase provides a basis for the development of new strategies to prevent the reperfusion-induced neuronal injury.

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Acknowledgements

This work was supported by cooperation Grant LA 1159/7-1 of the Deutsche Forschungsgemeinschaft (Germany) and Department of Science and Technology (India). This work was also supported by FAST-CoE grant from Ministry of HRD, Department of Education, Government of India, and was a part of the thesis of Megha Chagtoo supported by ICMR-SRF fellowship from Government of India and submitted in fulfillment of the requirements for the degree of Doctor of Philosophy at Integral University, Lucknow, India (IU/R&D/2017-MCN00060). We thank Lisa L. Schaffer (University of Tennessee) for the comprehensive editing of manuscript.

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  1. Madan M Godbole and Yury Ladilov contributed equally to this work.

Authors and Affiliations

  1. Department of Molecular Medicine and Biotechnology, Sanjay Gandhi Post Graduate Institute of Medical Sciences, Lucknow, India

    Megha Chagtoo, Swasti Tiwari & Madan M. Godbole

  2. Department of Bioscience, Integral University, Lucknow, India

    Megha Chagtoo & Neelam Pathak

  3. Department of Endocrine Surgery, Sanjay Gandhi Post Graduate Institute of Medical Sciences, Lucknow, India

    Nelson George

  4. Department of Clinical Pharmacology, Ruhr University Bochum, Bochum, Germany

    Yury Ladilov

  5. Center for Cardiovascular Research, Charité-Mitte, Hessische Strasse 3-4, 10115, Berlin, Germany

    Yury Ladilov

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  1. Megha Chagtoo
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Correspondence to Yury Ladilov.

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Chagtoo, M., George, N., Pathak, N. et al. Inhibition of Intracellular Type 10 Adenylyl Cyclase Protects Cortical Neurons Against Reperfusion-Induced Mitochondrial Injury and Apoptosis. Mol Neurobiol 55, 2471–2482 (2018). https://doi.org/10.1007/s12035-017-0473-y

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