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Quantitative In-Depth Profiling of the Postsynaptic Density Proteome to Understand the Molecular Mechanisms Governing Synaptic Physiology and Pathology

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Current Proteomic Approaches Applied to Brain Function

Part of the book series: Neuromethods ((NM,volume 127))

Abstract

The postsynaptic density (PSD) is a very large protein complex found in excitatory synapses, the most abundant synaptic type in the brain. PSDs contain key proteins to postsynaptic functioning, being closely related to synaptic plasticity and underlying high cognitive abilities such as learning and memory. Biochemical isolation of PSDs has allowed for their profiling by mass spectrometry-based proteomics, revealing it as a highly complex structure containing over 2000 proteins. Genetic studies have found many genes expressed at the PSD as causing neurological as well as mental and behavioral disorders. Understanding the proteomic architecture and dynamics of the PSD is not only key to understand normal brain function but also highly incapacitating human conditions. Here we present easy and robust protocols to isolate PSDs from very little amounts of brain tissue and a mass spectrometry proteomics procedure that allows to identify and quantify over 2000 different PSD proteins in a routinely fashion.

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Acknowledgments

This research was supported by Spanish grants BFU2012-34398 and BFU2015-69717-P, as well as AB personal fellowship Career Integration Grant, ref. 304111 (Marie-Curie Action) and Ramón y Cajal Fellowship (RYC-2011-08391). We thank Dr. Francesc Canals (Vall d’Hebron Institute of Oncology, Barcelona Spain) for critical review of mass spectrometry methods; Luna Martin (Vall d’Hebron Institute of Oncology, Barcelona Spain) for support on proteomic methods and analysis; Gemma Gou, Marcos Arranz, and Eva Olmedo (IIB Sant Pau, Barcelona Spain) for collaboration in experiments performance; and Ignasi Gich (IIB Sant Pau, Barcelona Spain) for support on biostatistics.

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Authors and Affiliations

  1. Molecular Physiology of the Synapse Laboratory, Biomedical Research Institute Sant Pau (IIB Sant Pau), St. Antoni Maria Claret St., 167, 08025, Barcelona, Spain

    Rita Reig-Viader & Àlex Bayés

  2. Universitat Autònoma de Barcelona, Cerdanyola del Vallès, Spain

    Rita Reig-Viader & Àlex Bayés

Authors
  1. Rita Reig-Viader
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  2. Àlex Bayés
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Correspondence to Àlex Bayés .

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Editors and Affiliations

  1. Clinical Neuroproteomics Unit, Navarrabiomed, Navarra Health Department, Public University of Navarra, Proteored-Institute of Health Carlos III (ISCIII), Navarra Institute for Health Research (IdiSNA), Pamplona, Spain

    Enrique Santamaría

  2. Clinical Neuroproteomics Unit, Navarrabiomed, Navarra Health Department, Public University of Navarra, Proteored-Institute of Health Carlos III (ISCIII), Navarra Institute for Health Research (IdiSNA), Pamplona, Spain

    Joaquín Fernández-Irigoyen

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Reig-Viader, R., Bayés, À. (2017). Quantitative In-Depth Profiling of the Postsynaptic Density Proteome to Understand the Molecular Mechanisms Governing Synaptic Physiology and Pathology. In: Santamaría, E., Fernández-Irigoyen, J. (eds) Current Proteomic Approaches Applied to Brain Function. Neuromethods, vol 127. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7119-0_16

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  • DOI: https://doi.org/10.1007/978-1-4939-7119-0_16

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7118-3

  • Online ISBN: 978-1-4939-7119-0

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