ArticleJournal of neuroscience methods2025
An immunohistochemical protocol for visualizing adrenergic receptor subtypes in the rhesus macaque hippocampus.
Article in Journal of neuroscience methods, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.
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Abstract
backgroundThe noradrenergic system is an important modulatory system in the brain, and dysfunction in this system is implicated in multiple neurodegenerative diseases. The study of this system in neuronal tissues relies on the availability of specific antibodies but to date no protocol exists for immunohistological visualization of α1, α2, and β adrenergic receptors in rhesus macaques. NEW
methodHere, we test the ability of various commercially available antibodies to detect these receptors in the primate brain and develop a protocol for visualization of receptors alongside noradrenergic axons and glial and vascular cells that interact with the noradrenergic system.
resultsOf the eleven primary antibodies for adrenergic receptors tested, five did not produce staining at any concentration. The remaining six antibodies underwent a preadsorption protocol to determine specificity of the antibody to its' immunogen sequence. Two antibodies failed this test, indicating they were binding to other targets in the brain. We then determined optimum concentrations for the remaining four antibodies. Additionally, we develop an immunofluorescence protocol that allows for the visualization of each AR - α1, α2a, or β1 - along with adrenergic axons as well as with glia and vasculature. COMPARISON WITH EXISTING
methodsWhile protocols exist for visualizing receptors in rodents, this is the first protocol for use in nonhuman primates.
conclusionsSeven out of the eleven tested antibodies were inaccurate, highlighting the importance of comprehensive testing. The stringent tests conducted here suggest that some commercially available antibodies can reliably detect adrenergic receptor subtypes in nonhuman primate tissue.
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