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Evaluation of the specificity of antibodies raised against cannabinoid receptor type 2 in the mouse retina

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Abstract

Cannabinoid receptors (CB1R and CB2R) are among the most abundant G protein-coupled receptors in the central nervous system. The endocannabinoid system is an attractive therapeutic target for immune system modulation and peripheral pain management. While CB1R is distributed in the nervous system, CB2R has traditionally been associated to the immune system. This dogma is currently a subject of debate since the discovery of CB2R expression in neurons using antibody-based methods. The localization of CB2R in the central nervous system (CNS) could have a significant impact on drug development because it would mean that in addition to its effects on the peripheral pain pathway, CB2R could also mediate some central effects of cannabinoids. In an attempt to clarify the debate over CB2R expression in the CNS, we tested several commercially or academically produced CB2R antibodies using Western blot and immunohistochemistry on retinal tissue obtained from wild-type mice and mice lacking CB2R (cnr2 −/−). One of the antibodies tested exhibited a valuable specificity as it marked a single band near the predicted molecular weight in Western blot and produced no staining in cnr2 −/− mice retina sections. The other antibodies tested detected multiple bands in Western blot and labeled unidentified proteins when used with their immunizing peptide or on cnr2 −/− retinal sections. We conclude that many commonly used antibodies raised against CB2R are not specific for use in immunohistochemistry, at least in the context of the mouse retina. Moreover, some of them tested presented significant lot-to-lot variability. Hence, caution should be used when interpreting prior and future studies using CB2R antibodies.

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Acknowledgments

We would like to thank Dr. Ken Mackie for kindly providing the CB2R antibodies and the constructive comments on this manuscript. This work was supported by a NSERC grant (194670–2009) to C.C., a CIHR grant (MOP 177796), and a NSERC grant (311892–2010) to J.-F.B. B.C. was supported by a Réseau Fonds de recherche Québec-Santé (FQRS) de recherche en santé de la vision studentship and J.-F.B. by a Chercheur-Boursier Junior 2 from the FRQS.

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Authors and Affiliations

  1. Laboratoire de Neuropharmacologie, École d’optométrie, Université de Montréal, C.P. 6128, Succursale Centre-Ville, Montreal, Québec, H3C 3J7, Canada

    Bruno Cécyre & Jean-François Bouchard

  2. Laboratoire des Neurosciences de la vision, École d’optométrie, Université de Montréal, Montreal, Québec, Canada

    Bruno Cécyre, Sébastien Thomas & Christian Casanova

  3. Chaire Harland Sanders, École d’optométrie, Université de Montréal, Montreal, Québec, Canada

    Maurice Ptito

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  1. Bruno Cécyre
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  2. Sébastien Thomas
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  3. Maurice Ptito
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  4. Christian Casanova
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  5. Jean-François Bouchard
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Correspondence to Jean-François Bouchard.

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Cécyre, B., Thomas, S., Ptito, M. et al. Evaluation of the specificity of antibodies raised against cannabinoid receptor type 2 in the mouse retina. Naunyn-Schmiedeberg's Arch Pharmacol 387, 175–184 (2014). https://doi.org/10.1007/s00210-013-0930-8

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