Evaluating the suitability of nicotinic acetylcholine receptor antibodies for standard immunodetection procedures.

Publisher: Wiley on behalf of the International Society for Neurochemistry Country of Publication: England NLM ID: 2985190R Publication Model: Print-Electronic Cited Medium: Print ISSN: 0022-3042 (Print) Linking ISSN: 00223042 NLM ISO Abbreviation: J Neurochem Subsets: MEDLINE

Publication: 2001- : Oxford, UK : Wiley on behalf of the International Society for Neurochemistry
Original Publication: New York : Raven Press

Antibodies/*metabolism ; Antibody Specificity/*immunology ; Immunohistochemistry/*methods ; Neurochemistry/*methods ; Protein Subunits/*immunology ; Receptors, Nicotinic/*immunology; Acetylcholine/metabolism ; Animals ; Animals, Newborn ; Antibodies/chemistry ; Blotting, Western ; Bungarotoxins/metabolism ; Cerebral Cortex/anatomy & histology ; Cerebral Cortex/immunology ; Cerebral Cortex/metabolism ; Electrophoresis, Gel, Two-Dimensional ; Female ; Hippocampus/anatomy & histology ; Hippocampus/immunology ; Hippocampus/metabolism ; Male ; Mice ; Mice, Inbred C57BL ; Mice, Knockout ; Neurons/immunology ; Neurons/metabolism ; Protein Subunits/analysis ; Protein Subunits/genetics ; Receptors, Nicotinic/analysis ; Receptors, Nicotinic/genetics ; Synaptic Transmission/immunology

Nicotinic acetylcholine receptors play important roles in numerous cognitive processes as well as in several debilitating central nervous system (CNS) disorders. In order to fully elucidate the diverse roles of nicotinic acetylcholine receptors in CNS function and dysfunction, a detailed knowledge of their cellular and subcellular localizations is essential. To date, methods to precisely localize nicotinic acetylcholine receptors in the CNS have predominantly relied on the use of anti-receptor subunit antibodies. Although data obtained by immunohistology and immunoblotting are generally in accordance with ligand binding studies, some discrepancies remain, in particular with electrophysiological findings. In this context, nicotinic acetylcholine receptor subunit-deficient mice should be ideal tools for testing the specificity of subunit-directed antibodies. Here, we used standard protocols for immunohistochemistry and western blotting to examine the antibodies raised against the alpha3-, alpha4-, alpha7-, beta2-, and beta4-nicotinic acetylcholine receptor subunits on brain tissues of the respective knock-out mice. Unexpectedly, for each of the antibodies tested, immunoreactivity was the same in wild-type and knock-out mice. These data imply that, under commonly used conditions, these antibodies are not suited for immunolocalization. Thus, particular caution should be exerted with regards to the experimental approach used to visualize nicotinic acetylcholine receptors in the brain.

0 (Antibodies)
0 (Bungarotoxins)
0 (Protein Subunits)
0 (Receptors, Nicotinic)
N9YNS0M02X (Acetylcholine)

Date Created: 20070411 Date Completed: 20070907 Latest Revision: 20220309

20221213

10.1111/j.1471-4159.2007.04498.x

17419810