The present address of N. Mechawar is the Douglas Hospital Research Centre, McGill University, Psychiatry, 6875 LaSalle Blvd., Verdun (Montréal), Québec, Canada H4H 1R3.
Evaluating the suitability of nicotinic acetylcholine receptor antibodies for standard immunodetection procedures
Article first published online: 20 FEB 2007
Journal of Neurochemistry
Volume 102, Issue 2, pages 479–492, July 2007
How to Cite
Moser, N., Mechawar, N., Jones, I., Gochberg-Sarver, A., Orr-Urtreger, A., Plomann, M., Salas, R., Molles, B., Marubio, L., Roth, U., Maskos, U., Winzer-Serhan, U., Bourgeois, J.-P., Le Sourd, A.-M., De Biasi, M., Schröder, H., Lindstrom, J., Maelicke, A., Changeux, J.-P. and Wevers, A. (2007), Evaluating the suitability of nicotinic acetylcholine receptor antibodies for standard immunodetection procedures. Journal of Neurochemistry, 102: 479–492. doi: 10.1111/j.1471-4159.2007.04498.x
- Issue published online: 20 FEB 2007
- Article first published online: 20 FEB 2007
- Received November 22, 2006; revised manuscript received January 15, 2007; accepted January 30, 2007.
- antibody specificity testing;
- knock-out mice;
- nicotinic acetylcholine receptor;
- western blotting
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 α3-, α4-, α7-, β2-, and β4-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.