Serotonin-induced down-regulation of cell surface serotonin transporter

Research output: Contribution to journal › Journal article › Research › peer-review

Standard

Serotonin-induced down-regulation of cell surface serotonin transporter. / Jørgensen, Trine Nygaard; Christensen, Peter Møller; Gether, Ulrik.

In: Neurochemistry International, Vol. 73, 07.2014, p. 107-12.

Research output: Contribution to journal › Journal article › Research › peer-review

Harvard

Jørgensen, TN, Christensen, PM & Gether, U 2014, 'Serotonin-induced down-regulation of cell surface serotonin transporter', Neurochemistry International, vol. 73, pp. 107-12. https://doi.org/10.1016/j.neuint.2014.01.005

APA

Jørgensen, T. N., Christensen, P. M., & Gether, U. (2014). Serotonin-induced down-regulation of cell surface serotonin transporter. Neurochemistry International, 73, 107-12. https://doi.org/10.1016/j.neuint.2014.01.005

Vancouver

Jørgensen TN, Christensen PM, Gether U. Serotonin-induced down-regulation of cell surface serotonin transporter. Neurochemistry International. 2014 Jul;73:107-12. https://doi.org/10.1016/j.neuint.2014.01.005

Author

Jørgensen, Trine Nygaard ; Christensen, Peter Møller ; Gether, Ulrik. / Serotonin-induced down-regulation of cell surface serotonin transporter. In: Neurochemistry International. 2014 ; Vol. 73. pp. 107-12.

Bibtex

@article{dbd51248c3cb4e8ba812a891a95aecf9,

title = "Serotonin-induced down-regulation of cell surface serotonin transporter",

abstract = "The serotonin transporter (SERT) terminates serotonergic signaling and enables refilling of synaptic vesicles by mediating reuptake of serotonin (5-HT) released into the synaptic cleft. The molecular and cellular mechanisms controlling SERT activity and surface expression are not fully understood. Here we demonstrate that the substrate 5-HT itself causes acute down-regulation of SERT cell surface expression. To assess surface SERT expression by ELISA, we used a SERT variant (TacSERT) where the N-terminus of SERT was fused to the intracellular tail of the extracellularly FLAG-tagged single-membrane spanning protein Tac. In stably transfected HEK293 cells, 5-HT caused a dose-dependent reduction in TacSERT surface signal with an EC50 value equivalent to the Km value observed for 5-HT uptake. The 5-HT-induced reduction in surface signal reached maximum within 40-60min and was blocked by the selective SERT inhibitor S-citalopram. 5-HT-induced reduction in SERT expression was further supported by surface biotinylation experiments showing 5-HT-induced reduction in wild type SERT plasma membrane levels. Moreover, preincubation with 5-HT lowered the Vmax for 5-HT uptake in cultured raphe serotonergic neurons, indicting that endogenous cell-surface resident SERT likewise is down-regulated in the presence of substrate.",

keywords = "Biotinylation, Down-Regulation, HEK293 Cells, Humans, Kinetics, Membrane Proteins, Neurons, Primary Cell Culture, Raphe Nuclei, Serotonin, Serotonin Plasma Membrane Transport Proteins, Transfection",

author = "J{\o}rgensen, {Trine Nygaard} and Christensen, {Peter M{\o}ller} and Ulrik Gether",

year = "2014",

month = jul,

doi = "10.1016/j.neuint.2014.01.005",

language = "English",

volume = "73",

pages = "107--12",

journal = "Neurochemistry International",

issn = "0197-0186",

publisher = "Elsevier",

}

RIS

TY - JOUR

T1 - Serotonin-induced down-regulation of cell surface serotonin transporter

AU - Jørgensen, Trine Nygaard

AU - Christensen, Peter Møller

AU - Gether, Ulrik

PY - 2014/7

Y1 - 2014/7

N2 - The serotonin transporter (SERT) terminates serotonergic signaling and enables refilling of synaptic vesicles by mediating reuptake of serotonin (5-HT) released into the synaptic cleft. The molecular and cellular mechanisms controlling SERT activity and surface expression are not fully understood. Here we demonstrate that the substrate 5-HT itself causes acute down-regulation of SERT cell surface expression. To assess surface SERT expression by ELISA, we used a SERT variant (TacSERT) where the N-terminus of SERT was fused to the intracellular tail of the extracellularly FLAG-tagged single-membrane spanning protein Tac. In stably transfected HEK293 cells, 5-HT caused a dose-dependent reduction in TacSERT surface signal with an EC50 value equivalent to the Km value observed for 5-HT uptake. The 5-HT-induced reduction in surface signal reached maximum within 40-60min and was blocked by the selective SERT inhibitor S-citalopram. 5-HT-induced reduction in SERT expression was further supported by surface biotinylation experiments showing 5-HT-induced reduction in wild type SERT plasma membrane levels. Moreover, preincubation with 5-HT lowered the Vmax for 5-HT uptake in cultured raphe serotonergic neurons, indicting that endogenous cell-surface resident SERT likewise is down-regulated in the presence of substrate.

AB - The serotonin transporter (SERT) terminates serotonergic signaling and enables refilling of synaptic vesicles by mediating reuptake of serotonin (5-HT) released into the synaptic cleft. The molecular and cellular mechanisms controlling SERT activity and surface expression are not fully understood. Here we demonstrate that the substrate 5-HT itself causes acute down-regulation of SERT cell surface expression. To assess surface SERT expression by ELISA, we used a SERT variant (TacSERT) where the N-terminus of SERT was fused to the intracellular tail of the extracellularly FLAG-tagged single-membrane spanning protein Tac. In stably transfected HEK293 cells, 5-HT caused a dose-dependent reduction in TacSERT surface signal with an EC50 value equivalent to the Km value observed for 5-HT uptake. The 5-HT-induced reduction in surface signal reached maximum within 40-60min and was blocked by the selective SERT inhibitor S-citalopram. 5-HT-induced reduction in SERT expression was further supported by surface biotinylation experiments showing 5-HT-induced reduction in wild type SERT plasma membrane levels. Moreover, preincubation with 5-HT lowered the Vmax for 5-HT uptake in cultured raphe serotonergic neurons, indicting that endogenous cell-surface resident SERT likewise is down-regulated in the presence of substrate.

KW - Biotinylation

KW - Down-Regulation

KW - HEK293 Cells

KW - Humans

KW - Kinetics

KW - Membrane Proteins

KW - Neurons

KW - Primary Cell Culture

KW - Raphe Nuclei

KW - Serotonin

KW - Serotonin Plasma Membrane Transport Proteins

KW - Transfection

U2 - 10.1016/j.neuint.2014.01.005

DO - 10.1016/j.neuint.2014.01.005

M3 - Journal article

C2 - 24462583

VL - 73

SP - 107

EP - 112

JO - Neurochemistry International

JF - Neurochemistry International

SN - 0197-0186

ER -

ID: 138813718