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Volume 52 (10): 1367-1376, 2004
Copyright ©The Histochemical Society, Inc.
Biochemical and Histochemical Evidence of Nonspecific Binding of 
7nAChR Antibodies to Mouse Brain Tissue
Alzheimer Research Laboratory, Department of Pharmacology and Therapeutics, University of South Florida, Tampa, Florida
Correspondence to: Marcia N. Gordon, University of South Florida, 12901 Bruce B. Downs Blvd., MDC Box 9, Tampa, FL 33612-4799. E-mail: mgordon{at}hsc.usf.edu
Alpha 7 nicotinic acetylcholine receptors are involved in learning and memory, and are implicated in the pathology of Alzheimer's disease and schizophrenia. Detection of 
7 subunits can be accomplished via immunodetection or -bungarotoxin-binding techniques. Standard protocols for immunohistochemistry and Western blotting were followed using several commercially available antibodies. Various mice were evaluated, including non-transgenics, APP, PS1, APP+PS1, and 7 knockouts. Initial results with amyloid-depositing mice revealed 7 immunolabeled astrocytes, in addition to expected neuronal staining. Subsequent studies with intrahippocampal injections of lipopolysaccharide (LPS) into 7 knockout mice showed that both neuronal and astrocytic labeling by 7 antibodies was nonspecific. On Western blots of mouse brain proteins, none of the bands detected with antibodies directed against 7 subunits diminished in the 7 knockout mice. Although LPS-related changes in the expression of some bands were found, these also were unaffected by the 7 genotype of the mice. In general, the Western staining patterns for these antibodies revealed few overlapping bands. These immunodetection data are in contrast to genotyping results and mRNA analyses that confirmed the disruption of the 7 allele and lack of 7 message in the knockouts. These findings suggest caution in interpreting results when using several commercially available 7 nicotinic receptor antibodies.
(J Histochem Cytochem 52:1367–1375, 2004)
Key Words: transgenic • lipopolysaccharide • alpha 7 nicotinic receptor • astrocytes • immunohistochemistry • Western blot
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