Abstract
A growing number of G protein-coupled receptors (GPCRs) have been identified on nuclear membranes. In many cases, it is unknown how the intracellular GPCR is activated, how it is trafficked to nuclear membranes, and what long-term signaling consequences follow nuclear receptor activation. Here we describe how to isolate nuclei that are free from plasma membrane and cytoplasmic contamination yet still exhibit physiological properties following receptor activation.
This is a preview of subscription content, log in via an institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsSpringer Nature is developing a new tool to find and evaluate Protocols. Learn more
References
Tadevosyan A, Vaniotis G, Allen BG et al (2012) G protein-coupled receptor signalling in the cardiac nuclear membrane: evidence and possible roles in physiological and pathophysiological function. J Physiol 590:1313–1330
Vaniotis G, Allen BG, Hébert TE (2011) Nuclear GPCRs in cardiomyocytes: an insider’s view of β-adrenergic receptor signaling. Am J Physiol Heart Circ Physiol 301:H1754–H1764
Bkaily G, Avedanian L, Al-Khoury J et al (2012) Receptors and ionic transporters in nuclear membranes: new targets for therapeutical pharmacological interventions. Can J Physiol Pharmacol 90:953–965
O’Malley KL, Jong YJ, Gonchar Y et al (2003) Activation of metabotropic glutamate receptor mGlu5 on nuclear membranes mediates intranuclear Ca2+ changes in heterologous cell types and neurons. J Biol Chem 278:28210–28219
Jong YJ, Kumar V, Kingston AE et al (2005) Functional metabotropic glutamate receptors on nuclei from brain and primary cultured striatal neurons. Role of transporters in delivering ligand. J Biol Chem 280:30469–30480
Isermann P, Lammerding J (2013) Nuclear mechanics and mechanotransduction in health and disease. Curr Biol 23:R1113–R1121
Rothballer A, Kutay U (2013) Poring over pores: nuclear pore complex insertion into the nuclear envelope. Trends Biochem Sci 38:292–301
Adams RL, Wente SR (2013) Uncovering nuclear pore complexity with innovation. Cell 152:1218–1221
Schreiber KH, Kennedy BK (2013) When lamins go bad: nuclear structure and disease. Cell 152:1365–1375
Kumar V, Jong YJ, O’Malley KL (2008) Activated nuclear metabotropic glutamate receptor mGlu5 couples to nuclear Gq/11 proteins to generate inositol 1,4,5-trisphosphate-mediated nuclear Ca2+ release. J Biol Chem 283:14072–14083
Jong YJ, Kumar V, O’Malley KL (2009) Intracellular metabotropic glutamate receptor 5 (mGluR5) activates signaling cascades distinct from cell surface counterparts. J Biol Chem 284:35827–35838
Kumar V, Fahey PG, Jong YJ et al (2012) Activation of intracellular metabotropic glutamate receptor 5 in striatal neurons leads to up-regulation of genes associated with sustained synaptic transmission including Arc/Arg3.1 protein. J Biol Chem 287:5412–5425
Acknowledgements
This work was supported by National Institutes of Health Grants MH57817 and MH69646 and NS081454. This work was also supported by a FRAXA Research Grant, by the Simons Foundation, and by the Bakewell Family Foundation.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2015 Springer Science+Business Media New York
About this protocol
Cite this protocol
Jong, YJ.I., O’Malley, K.L. (2015). Functional G Protein-Coupled Receptors on Nuclei from Brain and Primary Cultured Neurons. 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_10
Download citation
DOI: https://doi.org/10.1007/978-1-4939-1755-6_10
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-1754-9
Online ISBN: 978-1-4939-1755-6
eBook Packages: Springer Protocols