Journal of Molecular Histology

, Volume 38, Issue 4, pp 333–340 | Cite as

Endosomal location of dopamine receptors in neuronal cell cytoplasm

  • Elizabeth C. Wolstencroft
  • Goran Simic
  • Nguyen thi Man
  • Ian Holt
  • Le Thanh Lam
  • Paul R. Buckland
  • Glenn E. Morris
Original Paper

Abstract

Five subtypes of dopamine receptor exist in two subfamilies: two D1-like (D1 and D5) and three D2-like (D2, D3 and D4). We produced novel monoclonal antibodies against all three D2-like receptors and used them to localize receptors in Ntera-2 (NT-2) cells, the human neuronal precursor cell line. Most of the immunostaining for all three receptors colocalized with mannose-6-phosphate receptor, a marker for late endosomes formed by internalization of the plasma membrane. This result was obtained with antibodies against three different epitopes on the D3 receptor, to rule out the possibility of cross-reaction with another protein, and controls without primary antibody or in the presence of competitor antigen were completely negative. In rat cerebral cortex and hippocampus, some of the dopamine receptor staining was found in similar structures in neuronal cell cytoplasm. Only some of the neurons were positive for dopamine receptors and the pattern was consistent with previously-reported patterns of innervation by dopamine-producing neurons. Endosomal dopamine receptors may provide a useful method for identifying cell bodies of dopamine-responsive neurons to complement methods that detect only active receptors in the neuronal cell membrane.

Keywords

G protein-coupled receptor Endosome NT-2 cells Ntera-2 Monoclonal antibody Brain Hippocampus Cerebellum Cerebral cortex Mannose-6-phosphate receptor 

Notes

Acknowledgements

This study was supported by a studentship (Robert Hughes Jones Bequest for Research into Schizophrenia) from the Clwyd Institute of Health Studies for E.C.W. We thank Prof. P.W. Andrews (University of Sheffield, UK) for the Ntera-2 cell line. G.E.M. and G.S. thank the British Council in Croatia for support with travel grants.

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Copyright information

© Springer Science+Business Media B.V. 2007

Authors and Affiliations

  • Elizabeth C. Wolstencroft
    • 1
    • 5
  • Goran Simic
    • 3
  • Nguyen thi Man
    • 1
    • 2
  • Ian Holt
    • 1
    • 2
  • Le Thanh Lam
    • 1
    • 2
  • Paul R. Buckland
    • 4
  • Glenn E. Morris
    • 1
    • 2
  1. 1.Wolfson Centre for Inherited Neuromuscular DiseaseRJAH Orthopaedic HospitalOswestryUK
  2. 2.Institute of Science and Technology in MedicineKeele UniversityKeeleUK
  3. 3.Department of Neuroscience, Croatian Institute for Brain ResearchUniversity Medical SchoolZagrebCroatia
  4. 4.Department of Psychological MedicineUniversity of Wales College of MedicineCardiffUK
  5. 5.Royal Devon and Exeter NHS Foundation TrustExeterUK

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