Biophysical validation of serotonin 5-HT2A and 5-HT2C receptor interaction

PloS One
Daniel E FelsingKathryn A Cunningham

Abstract

The serotonin (5-HT) 5-HT2A receptor (5-HT2AR) and 5-HT2C receptor (5-HT2CR) in the central nervous system are implicated in a range of normal behaviors (e.g., appetite, sleep) and physiological functions (e.g., endocrine secretion) while dysfunctional 5-HT2AR and/or 5-HT2CR are implicated in neuropsychiatric disorders (e.g., addiction, obesity, schizophrenia). Preclinical studies suggest that the 5-HT2AR and 5-HT2CR may act in concert to regulate the neural bases for behavior. Here, we utilize three distinct biophysical and immunocytochemistry-based approaches to identify and study this receptor complex in cultured cells. Employing a split luciferase complementation assay (LCA), we demonstrated that formation of the 5-HT2AR:5-HT2CR complex exists within 50 nm, increases proportionally to the 5-HT2CR:5-HT2AR protein expression ratio, and is specific to the receptor interaction and not due to random complementation of the luciferase fragments. Using a proximity ligation assay (PLA), we found that cells stably expressing both the 5-HT2AR and 5-HT2CR exhibit 5-HT2AR:5-HT2CR heteroreceptor complexes within 40 nm of each other. Lastly, bioluminescence resonance energy transfer (BRET) analyses indicates the formation of a specific an...Continue Reading

References

Feb 1, 1990·Proceedings of the National Academy of Sciences of the United States of America·D JuliusT M Jessell
Jan 16, 1995·The Journal of Comparative Neurology·D E WrightL Jennes
Apr 1, 1994·Brain Research. Molecular Brain Research·M PompeianoG Mengod
Jan 14, 1999·Neuroscience Letters·A VysokanovD J Surmeier
Feb 3, 1999·Annals of the New York Academy of Sciences·K A BergW P Clarke
Mar 5, 2004·European Journal of Pharmaceutical Sciences : Official Journal of the European Federation for Pharmaceutical Sciences·Graeme Milligan
Jul 31, 2004·Proceedings of the National Academy of Sciences of the United States of America·Kathryn E LukerDavid Piwnica-Worms
May 18, 2007·European Journal of Pharmacology·Katharine Herrick-DavisBarbara A Weaver
Feb 26, 2008·Nature·Javier González-MaesoStuart C Sealfon
Jun 24, 2008·Behavioural Brain Research·Jason Hannon, Daniel Hoyer
Aug 5, 2008·Trends in Pharmacological Sciences·Mark J MillanClotilde Mannoury la Cour
Nov 10, 2009·Current Opinion in Pharmacology·Mohammed A Ayoub, Kevin D G Pfleger
Jun 8, 2011·Neuropharmacology·Laura AlbizuStuart C Sealfon
Aug 30, 2012·Nature Methods·Caroline A SchneiderKevin W Eliceiri
Dec 14, 2012·Nature·Jérémy BesnardAndrew L Hopkins
Jan 25, 2013·The Journal of Neuroscience : the Official Journal of the Society for Neuroscience·Noelle C AnastasioKathryn A Cunningham
Jul 16, 2013·Neuropharmacology·Kathryn A Cunningham, Noelle C Anastasio
Aug 14, 2013·Neuropsychopharmacology : Official Publication of the American College of Neuropsychopharmacology·Noelle C AnastasioKathryn A Cunningham
Dec 7, 2013·Biochemical and Biophysical Research Communications·Dasiel O Borroto-EscuelaKjell Fuxe
Feb 12, 2014·Pharmacological Reviews·Sergi FerréXavier Guitart
Nov 22, 2014·Postgraduate Medicine·Louis AronneSteven Smith
Dec 17, 2014·Pharmacological Reviews·Leonard L Howell, Kathryn A Cunningham
Jan 23, 2015·Molecular Pharmacology·Katharine Herrick-DavisJoseph E Mazurkiewicz
Mar 15, 2015·The Journal of Biological Chemistry·Maud KamalRalf Jockers
Jan 16, 2016·Disease Models & Mechanisms·Malgorzata S Martin-GronertSusan E Ozanne
Mar 5, 2017·The Journal of Biological Chemistry·Imane MoutkineStephane Doly
Oct 20, 2017·Expert Opinion on Drug Discovery·Zeyad T Sahli, Frank I Tarazi
Dec 1, 2017·BMC Bioinformatics·Curtis T RuedenKevin W Eliceiri
Mar 30, 2018·Nature Communications·Gemma NavarroSergi Ferré

❮ Previous
Next ❯

Citations

Sep 26, 2020·Frontiers in Integrative Neuroscience·Stefani StefaniSavvas S Papacostas
May 1, 2019·ACS Chemical Neuroscience·Barbara ChruścickaHarriët Schellekens
Jan 16, 2019·ACS Chemical Neuroscience·Amanda E PriceKathryn A Cunningham

❮ Previous
Next ❯

Methods Mentioned

BETA
PCR
BRET
proximity ligation assay
fluorescence imaging
transfection
saturation
protein assay
saturation binding
co-immunoprecipitation

Software Mentioned

Duolink Image Tool
Leica LASX
GraphPad
Leica Application Suite
FLIPR TETRA ScreenWorks
GraphPad Prism
NIH ImageJ

Related Concepts

Related Feeds

Addiction

This feed focuses mechanisms underlying addiction and addictive behaviour including heroin and opium dependence, alcohol intoxication, gambling, and tobacco addiction.