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The Journal of Neuroscience, November 1, 1999, 19(21):9281-9288

Receptor Subtype-Induced Targeting and Subtype-Specific Internalization of Human alpha 2-Adrenoceptors in PC12 Cells

Tuire Olli-Lähdesmäki1, 2, 3, Jaana Kallio1, 2, and Mika Scheinin1, 2

1 Department of Pharmacology and Clinical Pharmacology, 2 Medicity Research Laboratories, and 3 Turku Graduate School of Biomedical Sciences, University of Turku, FIN-20520 Turku, Finland

The three alpha 2-adrenergic receptor subtypes have distinct tissue distributions, desensitization properties, and, in some cell types, subtype-specific subcellular localization and trafficking properties. The subtypes also differ in their neuronal physiology. Therefore, we have investigated the localization and targeting of human alpha 2-adrenoceptors (alpha 2-AR) in PC12 cells, which were transfected to express the alpha 2-AR subtypes A, B, and C. Inspection of the receptors by indirect immunofluorescence and confocal microscopy showed that alpha 2A-AR were mainly targeted to the tips of the neurites, alpha 2B-AR were evenly distributed in the plasma membrane, and alpha 2C-AR were mostly located in an intracellular perinuclear compartment. After agonist treatment, alpha 2A- and alpha 2B-AR were internalized into partly overlapping populations of intracellular vesicles. Receptor subtype-specific changes in PC12 cell morphology were also discovered: expression of alpha 2A-AR, but not of alpha 2B- or alpha 2C-AR, induced differentiation-like changes in cells not treated with NGF. Also alpha 2B-AR were targeted to the tips of neurites when they were coexpressed in the same cells with alpha 2A-AR, indicating that the targeting of receptors to the tips of neurites is a consequence of a change in PC12 cell membrane protein trafficking that the alpha 2A-subtype induces. The marked agonist-induced internalization of alpha 2A-AR observed in both nondifferentiated and differentiated PC12 cells contrasts with earlier results from non-neuronal cells and points out the importance of the cellular environment for receptor endocytosis and trafficking. The targeting of alpha 2A-AR to nerve terminals in PC12 cells is in line with the putative physiological role of this receptor subtype as a presynaptic autoreceptor.

Key words: alpha 2-adrenoceptors; endocytosis; internalization; differentiation; PC12 cells; immunocytochemistry; targeting

Copyright © 1999 Society for Neuroscience  0270-6474/99/19219281-08$05.00/0


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