Co-immunoprecipitation of Membrane-Bound Receptors from Subsynaptic Compartments

  • Wilber Romero-FernandezEmail author
  • Maria Garcia-Barcelo
  • Yunis Perez-Betancourt
Part of the Neuromethods book series (NM, volume 144)


The allosteric modulation in G protein-coupled heteroreceptors have opened a new vision of the physiology and pathophysiology of the central nervous system. Understanding the role of the receptor-receptor interactions in the synaptic compartments is essential to elucidate the function of these macromolecular complexes in the brain intercellular communication that occurs within wiring and volume transmission. Here, we describe a procedure to identify the receptor-receptor interactions in different enriched subsynaptic fractions. First, we present a protocol to obtain synaptosomes by means of a discontinuous sucrose gradient. The method continues with a subcellular fractionation to separate the pre-synaptic, the post-synaptic, and the extra-synaptic fractions. Finally, we describe the Co-immunoprecipitation as an accepted technique to identify the receptor-receptor interaction from isolated and validated subsynaptic compartments.

Key words

G protein-coupled receptors Heteroreceptors Receptor-receptor interaction Synaptosome Sucrose gradient Subcellular fractionation Synaptic compartments Co-immunoprecipitation 



This work was supported by Prometeo Program of the Higher Education, Science, Technology and Innovation Secretary and the Technical University of Ambato (grants 1243-CU-P-2014 to WR-F, 0912-CU-P-2016 and 0932-CU-P-2016 to WR-F and YP-B, and 0906-CU-P-2016 to MG-B).


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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Wilber Romero-Fernandez
    • 1
    • 2
    Email author
  • Maria Garcia-Barcelo
    • 2
  • Yunis Perez-Betancourt
    • 3
  1. 1.Department of Cell and Molecular BiologyUppsala UniversityUppsalaSweden
  2. 2.Faculty of Health SciencesTechnical University of AmbatoAmbatoEcuador
  3. 3.Faculty of Food Science and EngineeringTechnical University of AmbatoAmbatoEcuador

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