Targeting of the 5-HT1A serotonin receptor to neuronal dendrites is mediated by Yif1B.
The 5-HT(1A) receptor (5-HT(1A)R) is the most extensively characterized serotonin (5-HT) receptor mainly because of its involvement in the mode of action of antidepressants. The 5-HT(1A)R is confined to the somatodendritic domain of central neurons, where it mediates serotonin-evoked hyperpolarization. Our previous studies underlined the role of the short 5-HT(1A)R C-terminal domain in receptor targeting to dendrites. We used this 17 aa region as bait in a yeast two-hybrid screen, and identified, for the first time, an intracellular protein interacting with the 5-HT(1A)R. This protein is homologous to the yeast Yif1p, previously implicated in vesicular trafficking between the endoplasmic reticulum (ER) and the Golgi apparatus, but not yet characterized in mammals. We confirmed 5-HT(1A)R-Yif1B interaction by glutathione S-transferase pull-down experiments using rat brain extracts and transfected cell lines. Yif1B is highly expressed in the brain, and specifically in raphe 5-HT(1A)R-expressing neurons. Colocalization of Yif1B and 5-HT(1A)R was observed in small vesicles involved in transient intracellular trafficking. Last, inhibition of endogenous expression of Yif1B in primary neuron cultures by small interfering RNA specifically prevented the addressing of 5-HT(1A)R to distal portions of the dendrites, without affecting other receptors, such as sst2A, P2X(2), and 5-HT(3A) receptors. Together, our results provide strong evidence that Yif1B is a member of the ER/Golgi trafficking machinery, which plays a key role in specific targeting of 5-HT(1A)R to the neuronal dendrites. This finding opens up new pathways for the study of 5-HT(1A)R regulation by partner proteins and for the development of novel antidepressant drugs.
Carrel
Damien
D
Faculté de Médecine Pierre et Marie Curie, Université Pierre et Marie Curie-Paris 6, Site Pitié-Salpêtrière, Institut Fédératif de Recherche 70 des Neurosciences, Unité Mixte de Recherche 677, F-75013 Paris, France.
Masson
Justine
J
Al Awabdh
Sana
S
Capra
Catherine Borg
CB
Lenkei
Zsolt
Z
Hamon
Michel
M
Emerit
Michel Boris
MB
Darmon
Michèle
M
D000818:Animals; D001921:Brain / Q000378:metabolism; D019556:COS Cells; D002522:Chlorocebus aethiops; D003712:Dendrites / Q000378:metabolism*; D005982:Glutathione Transferase / Q000378:metabolism; D018374:LLC-PK1 Cells; D008297:Male; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D009474:Neurons / Q000378:metabolism*; D021381:Protein Transport / Q000502:physiology; D051381:Rats; D017207:Rats, Sprague-Dawley; D044282:Receptor, Serotonin, 5-HT1A / Q000378:metabolism*; D012701:Serotonin / Q000378:metabolism; D013552:Swine; D014018:Tissue Distribution; D014162:Transfection; D020798:Two-Hybrid System Techniques; D033921:Vesicular Transport Proteins / Q000235:genetics / Q000378:metabolism / Q000502:physiology*
D009419:Nerve Tissue Proteins; D033921:Vesicular Transport Proteins; C531168:Yif1B protein, rat; D044282:Receptor, Serotonin, 5-HT1A; D012701:Serotonin; D005982:Glutathione Transferase