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High Resolution Imaging and Function of Nuclear G Protein-Coupled Receptors (GPCRs)

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Nuclear G-Protein Coupled Receptors

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1234))

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.

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Acknowledgements

The electron micrographs shown in this chapter were obtained at electron microscopy facility in the Department of Pharmacology & Therapeutics, McGill University. The facility currently has a Philips CM120 electron microscope equipped with a Gatan digital camera. This work was supported by grants from Canadian Institutes of Health Research (CIHR). VKB is a recipient of CIHR Systems Biology studentship award at McGill University, Montreal, Canada. We thank Mrs. Hendrika Fernandez, Mrs. Isabelle Lahaie, and Ms. Johanne Ouellette for their technical expertise.

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Authors and Affiliations

  1. Department of Pharmacology and Therapeutics, McGill University, Montreal, QC, Canada

    Vikrant K. Bhosle, Alfredo Ribeiro-da-Silva & Sylvain Chemtob

  2. CHU Sainte-Justine Hospital Research Centre, Montreal, QC, Canada

    Vikrant K. Bhosle & Jose Carlos Rivera

  3. Research Centre of Maisonneuve-Rosemont Hospital, Montreal, QC, Canada

    Vikrant K. Bhosle, Jose Carlos Rivera & Sylvain Chemtob

  4. Department of Pharmacology, Université de Sherbrooke, Sherbrooke, QC, Canada

    Fernand Gobeil Jr.

  5. Departments of Pediatrics, Ophthalmology and Pharmacology, CHU Sainte-Justine Hospital Research Centre, Montreal, QC, Canada, H3T 1C5

    Sylvain Chemtob

Authors
  1. Vikrant K. Bhosle
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  2. Fernand Gobeil Jr.
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  3. Jose Carlos Rivera
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  4. Alfredo Ribeiro-da-Silva
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  5. Sylvain Chemtob
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Correspondence to Sylvain Chemtob .

Editor information

Editors and Affiliations

  1. Montreal Heart Institute & Université de Montréal, Montreal, Québec, Canada

    Bruce G. Allen

  2. Pharmacology and Therapeutics, McGill University, Montreal, Québec, Canada

    Terence E. Hébert

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Bhosle, V.K., Gobeil, F., Rivera, J.C., Ribeiro-da-Silva, A., Chemtob, S. (2015). High Resolution Imaging and Function of Nuclear G Protein-Coupled Receptors (GPCRs). In: Allen, B., Hébert, T. (eds) Nuclear G-Protein Coupled Receptors. Methods in Molecular Biology, vol 1234. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1755-6_8

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  • DOI: https://doi.org/10.1007/978-1-4939-1755-6_8

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-1754-9

  • Online ISBN: 978-1-4939-1755-6

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