title: Characterization of Dahl salt-sensitive rats with genetic disruption of the A2B adenosine receptor gene: implications for A2B adenosine receptor signaling during hypertension.
abstract: The A(2B) 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 A(2B)AR 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 A(2B)AR 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 A(2B)AR. Our data indicate varying roles for A(2B)AR 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.
authors.affiliation: Department of Pharmacology and Toxicology, Medical College of Wisconsin, 8701 Watertown Plank Road, Milwaukee, WI, 53226, USA.|Cardiovascular Center, Medical College of Wisconsin, Milwaukee, WI, USA.
authors.forename: Shraddha
authors.initials: S
authors.lastname: Nayak
authors.affiliation: Department of Pharmacology and Toxicology, Medical College of Wisconsin, 8701 Watertown Plank Road, Milwaukee, WI, 53226, USA.
authors.forename: Md Abdul H
authors.initials: MA
authors.lastname: Khan
authors.affiliation: Department of Pharmacology and Toxicology, Medical College of Wisconsin, 8701 Watertown Plank Road, Milwaukee, WI, 53226, USA.|Cardiovascular Center, Medical College of Wisconsin, Milwaukee, WI, USA.
authors.forename: Tina C
authors.initials: TC
authors.lastname: Wan
authors.affiliation: Division of Developmental Immunology, La Jolla Institute for Allergy and Immunology, La Jolla, CA, USA.
authors.forename: Hong
authors.initials: H
authors.lastname: Pei
authors.affiliation: Division of Developmental Immunology, La Jolla Institute for Allergy and Immunology, La Jolla, CA, USA.
authors.forename: Joel
authors.initials: J
authors.lastname: Linden
authors.affiliation: Department of Physiology and Human Molecular Genetics Center, Medical College of Wisconsin, Milwaukee, WI, USA.
authors.forename: Melinda R
authors.initials: MR
authors.lastname: Dwinell
authors.affiliation: Cardiovascular Center, Medical College of Wisconsin, Milwaukee, WI, USA.|Department of Physiology and Human Molecular Genetics Center, Medical College of Wisconsin, Milwaukee, WI, USA.
authors.forename: Aron M
authors.initials: AM
authors.lastname: Geurts
authors.affiliation: Department of Pharmacology and Toxicology, Medical College of Wisconsin, 8701 Watertown Plank Road, Milwaukee, WI, 53226, USA.
authors.forename: John D
authors.initials: JD
authors.lastname: Imig
authors.affiliation: Department of Pharmacology and Toxicology, Medical College of Wisconsin, 8701 Watertown Plank Road, Milwaukee, WI, 53226, USA. jauchamp@mcw.edu.|Cardiovascular Center, Medical College of Wisconsin, Milwaukee, WI, USA. jauchamp@mcw.edu.
authors.forename: John A
authors.initials: JA
authors.lastname: Auchampach
mesh_terms: D058908:Adenosine A2 Receptor Agonists / Q000494:pharmacology; D000631:Aminopyridines / Q000494:pharmacology; D000818:Animals; D001794:Blood Pressure / Q000187:drug effects; D000242:Cyclic AMP / Q000378:metabolism; D016207:Cytokines / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism; D006973:Hypertension / Q000235:genetics* / Q000503:physiopathology*; D015850:Interleukin-6 / Q000378:metabolism; D009154:Mutation / Q000187:drug effects; D051381:Rats; D020303:Rats, Inbred Dahl; D043704:Receptor, Adenosine A2B / Q000235:genetics* / Q000378:metabolism; D015398:Signal Transduction / Q000235:genetics*
chemical_list: D058908:Adenosine A2 Receptor Agonists; D000631:Aminopyridines; C518875:BAY 60-6583; D016207:Cytokines; D015850:Interleukin-6; D043704:Receptor, Adenosine A2B; D000242:Cyclic AMP
keywords: Adenosine; Adenosine receptor; Angiotensin; Gene targeting; Hypertension; Metabolic disease