Distinct Subcellular Distribution of Delta-opioid Receptor Fused with Various Tags in PC12 Cells

Overview

Journal Neurochem Res

Specialties Chemistry
Neurology

Date 2008 Mar 28

PMID 18365312

Citations 25

Authors

Hai-Bo Wang

Ji-Song Guan

Lan Bao

Xu Zhang

Affiliations

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Abstract

In small dorsal root ganglion neurons, delta-opioid receptors (DORs) have been found to be mainly distributed in the cytoplasm and often associated with the membrane of large dense-core vesicles (LDCVs) that contain neuropeptides. To study the distribution of DORs under various physiological or pharmacological conditions, the receptors fused with different tags are constructed, transfected into cells or animals, and examined with microscopy. In this study, we show that DOR with different tags have distinct patterns of subcellular distribution in neuroendocrine cells, PC12 cells. Both immunostaining and vesicle fraction analysis showed that the native DORs expressed in PC12 cells were mainly associated with LDCVs. In transfected PC12 cells, DOR tagged with Myc or hemagglutinin exhibited LDCV localization. However, DOR fused with GFP at N- or C-terminus was found to be mainly localized on the cell surface, and mediated the function of DOR agonist. Therefore, the distribution of DOR fused with GFP differs from the native DORs. These results suggest that the subcellular distribution of the receptor could be better presented by the fused tag with smaller molecular size.

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