title: High resolution imaging and function of nuclear G protein-coupled receptors (GPCRs).
abstract: The traditional view of G protein-coupled receptors (GPCRs) being inactivated upon their internalization has been repeatedly challenged in recent years. GPCRs, in addition to forming the largest family of cell surface receptors, can also be found on intracellular membranes such as nuclear membranes. Since the first experimental evidence of GPCRs at the nucleus in the early 1990s, approximately 30 different GPCRs have been localized at the nucleus by independent research groups, including ours. In this chapter, we describe several techniques commonly used for immuno-detection of nuclear GPCRs focusing on subcellular fractionation of proteins based on their localization and transmission electron microscopy (TEM) using primary cultured cells as well as tissue sections. We also describe the use of confocal microscopy to study nuclear calcium currents, which can further affect downstream events such as gene transcription, nuclear envelope breakdown, or its reconstruction and nucleocytoplasmic protein transport.
authors.affiliation: Department of Pharmacology and Therapeutics, McGill University, Montreal, QC, Canada.
authors.forename: Vikrant K
authors.initials: VK
authors.lastname: Bhosle
authors.forename: Fernand
authors.initials: F
authors.lastname: Gobeil
authors.forename: Jose Carlos
authors.initials: JC
authors.lastname: Rivera
authors.forename: Alfredo
authors.initials: A
authors.lastname: Ribeiro-da-Silva
authors.forename: Sylvain
authors.initials: S
authors.lastname: Chemtob
mesh_terms: D021581:Active Transport, Cell Nucleus; D000818:Animals; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism* / Q000648:ultrastructure; D042783:Endothelial Cells; D057054:Molecular Imaging / Q000379:methods*; D061251:Primary Cell Culture; D018160:Receptors, Cytoplasmic and Nuclear / Q000378:metabolism*; D043562:Receptors, G-Protein-Coupled / Q000378:metabolism*; D015398:Signal Transduction; D013347:Subcellular Fractions
chemical_list: D018160:Receptors, Cytoplasmic and Nuclear; D043562:Receptors, G-Protein-Coupled