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By Mary Spaan

ISBN-10: 0521658748

ISBN-13: 9780521658744

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Nat Protoc 8:265–281 18. Sun Y, Day RN, Periasamy A (2011) Investigating protein-protein interactions in living cells using fluorescence lifetime imaging microscopy. Borroto-Escuela DO, Romero-Fernandez W, Garriga P, Ciruela F, Narvaez M, Tarakanov AO, Palkovits M, Agnati LF, Fuxe K (2013) G protein-coupled receptor heterodimerization in the brain. Methods Enzymol 521:281–294 20. Zacharias DA, Violin JD, Newton AC, Tsien RY (2002) Partitioning of lipid-modified monomeric GFPs into membrane microdomains of live cells.

These include (1) evidence for close proximity or physical association in native tissue, using approaches such as co-immunolocalization and co-immunoprecipitation, (2) specific functional properties for the heteromeric GPCR in native tissue, including changes in pharmacological properties, and (3) the use of knockout animals to provide information as to whether absence of one of the components of the GPCR heterocomplex affects signaling or behavioral properties of the other component. Some recent examples of GPCR heterocomplexes with functional properties in vivo in animal models include the Gq/11-coupled ghrelin GHSR1a receptor and the Gi/ocoupled dopamine D2 receptor (the GHSR1a-D2 receptor heterocomplex is expressed in hypothalamic neurons and involved in feeding behavior) [31], the Gq/11-coupled gastrin-releasing peptide (GRP) receptor and the Gi/o-coupled μ−opioid receptor isoform MOR1D (exons 1–3 and 8–9; the GRPR-MOR1D receptor heterocomplex is expressed in the spinal cord and involved in morphine-independent scratching) [32], the Gi/o-coupled melatonin receptors MT1 and MT2 (the MT1-MT2 receptor heterocomplex mediates the effect of melatonin on the light sensitivity of rod photoreceptors in mice) [33], the Gi/o-coupled dopamine D1 and D2 receptors (the D1-D2 receptor heterocomplex is expressed in the striatum and affects antidepressant-like effects in rats) [34], and the Gq/11-coupled serotonin 5-HT2A receptor and the Gi/o-coupled mGlu2 receptor (the 5-HT2A-mGlu2 receptor heterocomplex is expressed in frontal cortical neurons and exerts psychoactive-like behavioral effects) [35].

Methods Cell Biol 117:433–450 Yadav PN, Kroeze WK, Farrell MS et al (2011) Antagonist functional selectivity: 5-HT2A serotonin receptor antagonists differentially Techniques for the Study of GPCR Heteromerization in Living Cells and Animal Models 46. 47. 48. 49. 50. 51. 52. 53. 54. 55. 56. 57. 58. 59. 60. regulate 5-HT2A receptor protein level in vivo. J Pharmacol Exp Ther 339:99–105 Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of proteindye binding.

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Enhancing Fairness Through a Social Contract by Mary Spaan

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