Document Detail


Cell-type specific targeting of the alpha 2c-adrenoceptor. Evidence for the organization of receptor microdomains during neuronal differentiation of PC12 cells.
MedLine Citation:
PMID:  10906149     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
We have previously shown differences in the intracellular targeting of alpha2a (alpha(2A))- and alpha2c (alpha(2C))-adrenoreceptors expressed in the same cell line (von Zastrow, M., Link, R., Daunt, D. , Barsh, G., and Kobilka, B. (1993) J. Biol. Chem. 268, 763-766; Daunt, D. A., Hurt, C., Hein, L., Kallio, J., Feng, F., and Kobilka, B. K. (1997) Mol. Pharmacol. 51, 711-720). alpha(2A)-Adrenoreceptors reside primarily in the plasma membrane in HEK 293 cells, while co-expressed alpha(2C)-adrenoreceptors are found mainly in an intracellular compartment. Since alpha(2c)-adrenoreceptors are expressed primarily in the brain, we compared the intracellular targeting of alpha(2C)-adrenoreceptors in two neuroendocrine cell lines with the targeting in three epithelial cell lines and one fibroblast cell line. In transiently transfected COS7 cells, and in stably transfected normal rat kidney cells, Madin-Darby canine kidney cells, and Rat1 fibroblasts, a significant proportion of alpha(2C)-adrenoreceptor detected by immunocytochemistry co-localized with markers for both the endoplasmic reticulum and the cis/medial Golgi compartments. In contrast, both PC12 cells and AtT20 cells efficiently targeted alpha(2C)-adrenoreceptors to the plasma membrane. Ligand binding and Western blot analyses indicate that intracellular receptor in normal rat kidney cells is functional and undergoes normal post-translational processing. In PC12 cells the expressed alpha(2C)-adrenoreceptors become concentrated in neurite outgrowths in discrete regions of the plasma membrane having a high density of F-actin following treatment with nerve growth factor. These findings provide evidence for cell-type specific factors that facilitate the targeting of the G protein-coupled receptors to the plasma membrane.
Authors:
C M Hurt; F Y Feng; B Kobilka
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Publication Detail:
Type:  Comparative Study; Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  The Journal of biological chemistry     Volume:  275     ISSN:  0021-9258     ISO Abbreviation:  J. Biol. Chem.     Publication Date:  2000 Nov 
Date Detail:
Created Date:  2000-11-27     Completed Date:  2001-01-04     Revised Date:  2007-11-14    
Medline Journal Info:
Nlm Unique ID:  2985121R     Medline TA:  J Biol Chem     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  35424-31     Citation Subset:  IM    
Affiliation:
Howard Hughes Medical Institute, Stanford University Medical School, Stanford, California 94305-5428, USA.
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MeSH Terms
Descriptor/Qualifier:
Animals
Blotting, Western
Brain / metabolism
COS Cells
Cell Differentiation
Cell Membrane / metabolism
Cyclic AMP / metabolism
Dexmedetomidine / pharmacology
Dogs
Dose-Response Relationship, Drug
Endoplasmic Reticulum / metabolism
Epitopes
Fibroblasts / metabolism
Fluorescent Antibody Technique, Indirect
Golgi Apparatus
Humans
Immunohistochemistry
Kidney / metabolism
Ligands
Membrane Microdomains / metabolism*
Nerve Growth Factor / pharmacology
Neurons / metabolism*
PC12 Cells
Protein Binding
Protein Structure, Tertiary
Rats
Receptors, Adrenergic, alpha-2 / metabolism*
Receptors, Cell Surface / metabolism
Transfection
Grant Support
ID/Acronym/Agency:
G07365//PHS HHS
Chemical
Reg. No./Substance:
0/Epitopes; 0/Ligands; 0/Receptors, Adrenergic, alpha-2; 0/Receptors, Cell Surface; 0/adrenergic receptor alpha(2C); 113775-47-6/Dexmedetomidine; 60-92-4/Cyclic AMP; 9061-61-4/Nerve Growth Factor

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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