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Detecting GPCR Complexes in Postmortem Human Brain with Proximity Ligation Assay and A Bayesian Classifier

Ying Zhu, József Mészáros, Roman Walle, Rongxi Fan, Ziyi Sun, Andrew J. Dwork, Pierre Trifilieff, Jonathan A. Javitch
doi: https://doi.org/10.1101/687780
Ying Zhu
1Division of Molecular Therapeutics, New York State Psychiatric Institute, New York, New York 10032, USA
2Department of Psychiatry, Columbia University, New York, New York 10032, USA
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József Mészáros
3Zuckerman Institute, Columbia University, New York, NY 10027, USA
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Roman Walle
4Univ. Bordeaux, INRA, Bordeaux INP, NutriNeuro, UMR 1286, F-33000, Bordeaux, France
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Rongxi Fan
1Division of Molecular Therapeutics, New York State Psychiatric Institute, New York, New York 10032, USA
2Department of Psychiatry, Columbia University, New York, New York 10032, USA
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Ziyi Sun
1Division of Molecular Therapeutics, New York State Psychiatric Institute, New York, New York 10032, USA
2Department of Psychiatry, Columbia University, New York, New York 10032, USA
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Andrew J. Dwork
2Department of Psychiatry, Columbia University, New York, New York 10032, USA
5Department of Pathology and Cell Biology, Columbia University, New York State Psychiatric Institute, New York, New York
6Division of Molecular Imaging and Neuropathology, New York State Psychiatric Institute, New York, New York
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Pierre Trifilieff
4Univ. Bordeaux, INRA, Bordeaux INP, NutriNeuro, UMR 1286, F-33000, Bordeaux, France
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Jonathan A. Javitch
1Division of Molecular Therapeutics, New York State Psychiatric Institute, New York, New York 10032, USA
2Department of Psychiatry, Columbia University, New York, New York 10032, USA
8Department of Pharmacology, Columbia University, New York, New York 10032, USA
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  • For correspondence: jonathan.javitch@nyspi.columbia.edu
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Abstract

Despite the general controversy regarding the existence and physiological relevance of Class A GPCR dimerization, there is substantial evidence for functional interactions between dopamine D2 receptor (D2R) and adenosine A2A receptor (A2AR). A2AR-D2R complexes have been detected in rodent brains by proximity ligation assay (PLA), but their existence in the human brain is yet to be demonstrated. In this study, we used brightfield PLA, combined with a systematic sampling and a parameter-free naïve Bayesian classifier, and demonstrated proximity between D2R and A2AR in the adult human ventral striatum, consistent with their colocalization within complexes and the possible existence of D2R-A2AR heteromers. These methods are applicable to the quantitative analysis of proximity of two proteins and the expression of individual proteins.

Method Summary Brightfield proximity ligation assay (PLA) was used to assess the expression of G protein-coupled receptors and their proximity in postmortem adult human brains. A novel automated machine learning method (Bayesian Optimized PLA Signal Sorting) was developed to automatically quantify brightfield PLA data.

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Posted July 02, 2019.
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Detecting GPCR Complexes in Postmortem Human Brain with Proximity Ligation Assay and A Bayesian Classifier
Ying Zhu, József Mészáros, Roman Walle, Rongxi Fan, Ziyi Sun, Andrew J. Dwork, Pierre Trifilieff, Jonathan A. Javitch
bioRxiv 687780; doi: https://doi.org/10.1101/687780
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Detecting GPCR Complexes in Postmortem Human Brain with Proximity Ligation Assay and A Bayesian Classifier
Ying Zhu, József Mészáros, Roman Walle, Rongxi Fan, Ziyi Sun, Andrew J. Dwork, Pierre Trifilieff, Jonathan A. Javitch
bioRxiv 687780; doi: https://doi.org/10.1101/687780

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