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Immunohistochemical localization of adenosine A 2A receptors in the rat central nervous system
Author(s) -
Rosin Diane L.,
Robeva Anna,
Woodard Robin L.,
Guyenet Patrice G.,
Linden Joel
Publication year - 1998
Publication title -
journal of comparative neurology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.855
H-Index - 209
eISSN - 1096-9861
pISSN - 0021-9967
DOI - 10.1002/(sici)1096-9861(19981116)401:2<163::aid-cne2>3.0.co;2-d
Subject(s) - nucleus accumbens , striatum , adenosine a2a receptor , globus pallidus , biology , neuropil , receptor , medicine , chemistry , neuroscience , central nervous system , basal ganglia , agonist , dopamine , adenosine receptor , biochemistry
The A 2A adenosine receptor (A 2A ‐AR) transcript and radioligand binding sites have a distinct distribution in rat brain, restricted primarily to the striatum, nucleus accumbens and olfactory tubercles. We describe here the use of purified recombinant human A 2A ‐ARs to generate a monoclonal antibody that has been used to better resolve the distribution of A 2A ‐ARs in rat brain. The antibody can detect 1 ng of purified recombinant receptor by Western blotting and is potent (EC 50 = 0.62 μg/ ml) and highly selective for the A 2A ‐AR subtype. By Western blotting, the apparent molecular mass of recombinant and rat striatal receptors shifts upon deglycosylation from 43–48 to 42 kilodaltons. Analyses of chimeric A 1 / A 2A ‐ARs and synthesis of a blocking peptide pinpointed the epitope (SQPLPGER) of the antibody to the center of the third intracellular loop of the receptor. Incubation of rat striatal membranes with antibody reduces receptor coupling to G‐proteins. In rat brain, dense A 2A ‐AR‐like immunoreactivity that is eliminated by the blocking peptide was found in the neuropil of the striatum, nucleus accumbens (rostral pole, core and shell), cell bridges of the striatum, olfactory tubercles, and areas of extended amygdala with somewhat lighter labeling in the globus pallidus and nucleus of the solitary tract. Light perikaryal labeling was found in other areas of the brain, including the cortex, hippocampus, thalamus, cerebellum, and portions of the hindbrain. The observed distribution of A 2A ‐AR immunoreactivity throughout the neuraxis is consistent with the receptors' role in modulating dopaminergic neurotransmission and central control of cardiovascular function. J. Comp. Neurol. 401:163–186, 1998. © 1998 Wiley‐Liss, Inc.