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The Journal of Neuroscience, December 15, 2000, 20(24):9111-9118

Dominant Role of the Cytosolic C-Terminal Domain of the Rat 5-HT_1B Receptor in Axonal-Apical Targeting

Nicolas Jolimay, Louis Franck, Xavier Langlois, Michel Hamon, and Michèle Darmon

Institut National de la Santé et de la Recherche Médicale U288, Faculté de Médecine Pitié-Salpêtrière, 75013 Paris, France

The 5-HT_1A and 5-HT_1B receptors for serotonin exhibit a different membrane localization to either soma and dendrites (5-HT_1AR) or axons and terminals (5-HT_1BR) of neurons in the CNS. The mechanisms responsible for their differential targeting were investigated previously by transfecting various 5-HT_1AR/5-HT_1BR chimeras in the epithelial Lilly pork kidney (LLC-PK1) cell line. This first study suggested that a specific targeting signal is located in the C-terminal portion (comprising the last two transmembrane and the cytoplasmic C-terminal domains) of the 5-HT_1Aand/or 5-HT_1B receptors. In the present study, the role of the cytosolic C-terminal tail of the receptors was further investigated by transfecting truncated receptors and 5-HT_1AR/5-HT_1BR chimeras in both the epithelial LLC-PK1 cells and rat hippocampal neurons in primary culture. Confocal microscopic analysis of immunofluorescence with specific anti-5-HTR antibodies and anti-microtubule-associated protein 2 or anti-neurofilament 200k antibodies showed that substitution of the cytosolic C-terminal tail of the 5-HT_1BR in the 5-HT_1AR addressed the resulting chimera to the axon of neurons and to the apical domain of LLC-PK1 cells. Therefore, the short tail of the 5-HT_1BR presents an apical targeting signal that can also act as an axonal targeting signal. In addition, a domain within the third intracytoplasmic loop of the 5-HT_1BR, responsible for its Golgi sequestration in LLC-PK1 cells, appeared to act as another axonal targeting signal in hippocampal neurons.

Key words: cell polarity; addressing mechanisms; axons; dendrites; serotonin receptors; 5-HT_1AR; 5-HT_1BR

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Copyright © 2000 Society for Neuroscience  0270-6474/00/20249111-08$05.00/0

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