The PDZ-binding domain is essential for the dendritic targeting of 5-HT2A serotonin receptors in cortical pyramidal neurons in vitro

Z. Xia, S. J. Hufeisen, John Gray, B. L. Roth

Research output: Contribution to journalArticle

48 Citations (Scopus)

Abstract

The 5-HT2A serotonin receptor represents an important molecular target for atypical antipsychotic drugs and for most hallucinogens. In the mammalian cerebral cortex, 5-HT2A receptors are enriched in pyramidal neurons, within which 5-HT2A receptors are preferentially sorted to the apical dendrites. In primary cortical cultures, 5-HT2A receptors are sorted to dendrites and not found in the axons of pyramidal neurons. We identified a sorting motif that mediates the preferential targeting of 5-HT2A receptors to the dendrites of cortical pyramidal neurons in vitro. We constructed green fluorescent protein-tagged 5-HT2A receptors wherein potential sorting motifs were disrupted, and subsequently employed either the Semliki Forest virus or calcium phosphate for the transient expression of recombinant 5-HT2A receptors in cultured cortical pyramidal neurons. Using dual-labeling immunofluorescent confocal microscopy, we quantified the axonal and dendritic sorting patterns of endogenous and recombinant 5-HT2A receptors. We discovered that disruption of the PDZ-binding domain of the 5-HT2A receptor greatly attenuates the dendritic targeting of 5-HT2A receptors without inappropriately sorting 5-HT2A receptors to axons. The PDZ-binding domain is therefore a necessary signal for the preferential targeting of the 5-HT 2A receptor to the dendritic compartment of cultured cortical pyramidal neurons, the first such role ascribed to this protein-protein interaction motif of any G protein-coupled receptor.

Original languageEnglish (US)
Pages (from-to)907-920
Number of pages14
JournalNeuroscience
Volume122
Issue number4
DOIs
StatePublished - 2003
Externally publishedYes

Fingerprint

PDZ Domains
Receptor, Serotonin, 5-HT2A
Pyramidal Cells
Dendrites
Axons
In Vitro Techniques
Protein Interaction Domains and Motifs
Semliki forest virus
Hallucinogens
G-Protein-Coupled Receptors
Green Fluorescent Proteins
Confocal Microscopy
Cerebral Cortex
Antipsychotic Agents

Keywords

  • Axon
  • Dendrite
  • G protein-coupled receptor
  • GPCR
  • Neuronal polarity
  • Polarized protein sorting

ASJC Scopus subject areas

  • Neuroscience(all)

Cite this

The PDZ-binding domain is essential for the dendritic targeting of 5-HT2A serotonin receptors in cortical pyramidal neurons in vitro. / Xia, Z.; Hufeisen, S. J.; Gray, John; Roth, B. L.

In: Neuroscience, Vol. 122, No. 4, 2003, p. 907-920.

Research output: Contribution to journalArticle

@article{2ff5a1fbef4c4bf2984dd856ad2423a1,
title = "The PDZ-binding domain is essential for the dendritic targeting of 5-HT2A serotonin receptors in cortical pyramidal neurons in vitro",
abstract = "The 5-HT2A serotonin receptor represents an important molecular target for atypical antipsychotic drugs and for most hallucinogens. In the mammalian cerebral cortex, 5-HT2A receptors are enriched in pyramidal neurons, within which 5-HT2A receptors are preferentially sorted to the apical dendrites. In primary cortical cultures, 5-HT2A receptors are sorted to dendrites and not found in the axons of pyramidal neurons. We identified a sorting motif that mediates the preferential targeting of 5-HT2A receptors to the dendrites of cortical pyramidal neurons in vitro. We constructed green fluorescent protein-tagged 5-HT2A receptors wherein potential sorting motifs were disrupted, and subsequently employed either the Semliki Forest virus or calcium phosphate for the transient expression of recombinant 5-HT2A receptors in cultured cortical pyramidal neurons. Using dual-labeling immunofluorescent confocal microscopy, we quantified the axonal and dendritic sorting patterns of endogenous and recombinant 5-HT2A receptors. We discovered that disruption of the PDZ-binding domain of the 5-HT2A receptor greatly attenuates the dendritic targeting of 5-HT2A receptors without inappropriately sorting 5-HT2A receptors to axons. The PDZ-binding domain is therefore a necessary signal for the preferential targeting of the 5-HT 2A receptor to the dendritic compartment of cultured cortical pyramidal neurons, the first such role ascribed to this protein-protein interaction motif of any G protein-coupled receptor.",
keywords = "Axon, Dendrite, G protein-coupled receptor, GPCR, Neuronal polarity, Polarized protein sorting",
author = "Z. Xia and Hufeisen, {S. J.} and John Gray and Roth, {B. L.}",
year = "2003",
doi = "10.1016/S0306-4522(03)00589-X",
language = "English (US)",
volume = "122",
pages = "907--920",
journal = "Neuroscience",
issn = "0306-4522",
publisher = "Elsevier Limited",
number = "4",

}

TY - JOUR

T1 - The PDZ-binding domain is essential for the dendritic targeting of 5-HT2A serotonin receptors in cortical pyramidal neurons in vitro

AU - Xia, Z.

AU - Hufeisen, S. J.

AU - Gray, John

AU - Roth, B. L.

PY - 2003

Y1 - 2003

N2 - The 5-HT2A serotonin receptor represents an important molecular target for atypical antipsychotic drugs and for most hallucinogens. In the mammalian cerebral cortex, 5-HT2A receptors are enriched in pyramidal neurons, within which 5-HT2A receptors are preferentially sorted to the apical dendrites. In primary cortical cultures, 5-HT2A receptors are sorted to dendrites and not found in the axons of pyramidal neurons. We identified a sorting motif that mediates the preferential targeting of 5-HT2A receptors to the dendrites of cortical pyramidal neurons in vitro. We constructed green fluorescent protein-tagged 5-HT2A receptors wherein potential sorting motifs were disrupted, and subsequently employed either the Semliki Forest virus or calcium phosphate for the transient expression of recombinant 5-HT2A receptors in cultured cortical pyramidal neurons. Using dual-labeling immunofluorescent confocal microscopy, we quantified the axonal and dendritic sorting patterns of endogenous and recombinant 5-HT2A receptors. We discovered that disruption of the PDZ-binding domain of the 5-HT2A receptor greatly attenuates the dendritic targeting of 5-HT2A receptors without inappropriately sorting 5-HT2A receptors to axons. The PDZ-binding domain is therefore a necessary signal for the preferential targeting of the 5-HT 2A receptor to the dendritic compartment of cultured cortical pyramidal neurons, the first such role ascribed to this protein-protein interaction motif of any G protein-coupled receptor.

AB - The 5-HT2A serotonin receptor represents an important molecular target for atypical antipsychotic drugs and for most hallucinogens. In the mammalian cerebral cortex, 5-HT2A receptors are enriched in pyramidal neurons, within which 5-HT2A receptors are preferentially sorted to the apical dendrites. In primary cortical cultures, 5-HT2A receptors are sorted to dendrites and not found in the axons of pyramidal neurons. We identified a sorting motif that mediates the preferential targeting of 5-HT2A receptors to the dendrites of cortical pyramidal neurons in vitro. We constructed green fluorescent protein-tagged 5-HT2A receptors wherein potential sorting motifs were disrupted, and subsequently employed either the Semliki Forest virus or calcium phosphate for the transient expression of recombinant 5-HT2A receptors in cultured cortical pyramidal neurons. Using dual-labeling immunofluorescent confocal microscopy, we quantified the axonal and dendritic sorting patterns of endogenous and recombinant 5-HT2A receptors. We discovered that disruption of the PDZ-binding domain of the 5-HT2A receptor greatly attenuates the dendritic targeting of 5-HT2A receptors without inappropriately sorting 5-HT2A receptors to axons. The PDZ-binding domain is therefore a necessary signal for the preferential targeting of the 5-HT 2A receptor to the dendritic compartment of cultured cortical pyramidal neurons, the first such role ascribed to this protein-protein interaction motif of any G protein-coupled receptor.

KW - Axon

KW - Dendrite

KW - G protein-coupled receptor

KW - GPCR

KW - Neuronal polarity

KW - Polarized protein sorting

UR - http://www.scopus.com/inward/record.url?scp=0344441294&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0344441294&partnerID=8YFLogxK

U2 - 10.1016/S0306-4522(03)00589-X

DO - 10.1016/S0306-4522(03)00589-X

M3 - Article

VL - 122

SP - 907

EP - 920

JO - Neuroscience

JF - Neuroscience

SN - 0306-4522

IS - 4

ER -