Regresar

Co-immunoprecipitation of Membrane-Bound Receptors from Subsynaptic Compartments

Abstract:

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.