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Characterization of Dahl salt-sensitive rats with genetic disruption of the A2B adenosine receptor gene: implications for A2B adenosine receptor signaling during hypertension

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Abstract

The A2B adenosine receptor (AR) has emerged as a unique member of the AR family with contrasting roles during acute and chronic disease states. We utilized zinc-finger nuclease technology to create A2BAR gene (Adora2b)-disrupted rats on the Dahl salt-sensitive (SS) genetic background. This strategy yielded a rat strain (SS-Adora2b mutant rats) with a 162-base pair in-frame deletion of Adora2b that included the start codon. Disruption of A2BAR function in SS-Adora2b mutant rats was confirmed by loss of agonist (BAY 60-6583 or NECA)-induced cAMP accumulation and loss of interleukin-6 release from isolated fibroblasts. In addition, BAY 60-6583 produced a dose-dependent increase in glucose mobilization that was absent in SS-Adora2b mutants. Upon initial characterization, SS-Adora2b mutant rats were found to exhibit increased body weight, a transient delay in glucose clearance, and reduced proinflammatory cytokine production following challenge with lipopolysaccharide (LPS). In addition, blood pressure was elevated to a greater extent (∼15–20 mmHg) in SS-Adora2b mutants as they aged from 7 to 21 weeks. In contrast, hypertension augmented by Ang II infusion was attenuated in SS-Adora2b mutant rats. Despite differences in blood pressure, indices of renal and cardiac injury were similar in SS-Adora2b mutants during Ang II-augmented hypertension. We have successfully created and validated a new animal model that will be valuable for investigating the biology of the A2BAR. Our data indicate varying roles for A2BAR signaling in regulating blood pressure in SS rats, playing both anti- and prohypertensive roles depending on the pathogenic mechanisms that contribute to blood pressure elevation.

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Acknowledgments

This study was supported by grants from the National Institutes of Health (HL077707, HL111392, and RC2 HL101681).

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Authors and Affiliations

  1. Department of Pharmacology and Toxicology, Medical College of Wisconsin, 8701 Watertown Plank Road, Milwaukee, WI, 53226, USA

    Shraddha Nayak, Md. Abdul H. Khan, Tina C. Wan, John D. Imig & John A. Auchampach

  2. Cardiovascular Center, Medical College of Wisconsin, Milwaukee, WI, USA

    Shraddha Nayak, Tina C. Wan, Aron M. Geurts & John A. Auchampach

  3. Division of Developmental Immunology, La Jolla Institute for Allergy and Immunology, La Jolla, CA, USA

    Hong Pei & Joel Linden

  4. Department of Physiology and Human Molecular Genetics Center, Medical College of Wisconsin, Milwaukee, WI, USA

    Melinda R. Dwinell & Aron M. Geurts

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  1. Shraddha Nayak
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Correspondence to John A. Auchampach.

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Nayak, S., Khan, M.A.H., Wan, T.C. et al. Characterization of Dahl salt-sensitive rats with genetic disruption of the A2B adenosine receptor gene: implications for A2B adenosine receptor signaling during hypertension. Purinergic Signalling 11, 519–531 (2015). https://doi.org/10.1007/s11302-015-9470-7

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