Immunohistochemical detection of A1 adenosine receptors in rat brain with emphasis on localization in the hippocampal formation, cerebral cortex, cerebellum, and basal ganglia.
Polyclonal antisera were generated against two identical regions of rat and human A1 adenosine receptors using synthetic multiple-antigenic-peptides as immunogens. Western blotting showed that the antisera recognized a single protein in brain of the expected size for A1 receptors. Immunohistochemistry of CHO cells transfected with the rat or human A1 adenosine receptor cDNAs showed robust labeling of the cell surface. In contrast, labeling was not apparent over non-transfected CHO cells, nor over CHO cells expressing A2a receptors. The pattern of immunoreactivity in rat brain was similar to that expected for A1 adenosine receptors. In contrast to receptor autoradiography or in situ hybridization methods, immunohistochemistry allowed identification of individually labeled cells and processes. Heavy labeling was apparent in many brain regions. In the hippocampal formation, strong labeling was present on granule cell bodies and dendrites, mossy fibers, and pyramidal neurons. In cerebellum, basket cells were the most heavily labeled cell type. Less intense staining was present over granule cells. In cerebral cortex, pyramidal cells were the most heavily labeled cell type, and some interneurons were also labeled. In the basal ganglia, 43% of neurons in the globus pallidus were labeled. In the caudate-putamen region, 38% of neurons were labeled. Heavy labeling was present in most thalamic nuclei, and moderate to heavy labeling was seen in many brainstem nuclei. These data identify specific cellular sites of A1 receptor expression and support the concept of cellular specificity of A1 adenosine receptor action.
Rivkees
S A
SA
Department of Pediatrics, Herman B Wells Center for Pediatric Research, James Whitcomb Riley Hospital for Children, Indianapolis, IN, USA.
Price
S L
SL
Zhou
F C
FC
D000818:Animals; D001479:Basal Ganglia / Q000737:chemistry; D015153:Blotting, Western; D001921:Brain / Q000033:anatomy & histology*; D001923:Brain Chemistry; D016466:CHO Cells / Q000166:cytology; D002531:Cerebellum / Q000737:chemistry; D002540:Cerebral Cortex / Q000737:chemistry; D006224:Cricetinae; D006624:Hippocampus / Q000737:chemistry; D007106:Immune Sera; D007150:Immunohistochemistry; D008297:Male; D011817:Rabbits; D051381:Rats; D017208:Rats, Wistar; D018047:Receptors, Purinergic P1 / Q000032:analysis*
D007106:Immune Sera; D018047:Receptors, Purinergic P1