Neurotrophin-regulated sorting of opioid receptors in the biosynthetic pathway of neurosecretory cells.
Neurotrophins modulate the endogenous opioid system, but the underlying mechanisms are poorly understood. We observed an unexpected effect of neurotrophin signaling on the membrane trafficking of recombinant opioid receptors expressed in neurosecretory cells. Epitope-tagged delta opioid receptor (DOR) and mu opioid receptor (MOR) were differentially localized between surface and internal membrane pools, respectively, when expressed in primary cultured hippocampal neurons, consistent with previous studies by others of natively expressing neurons. Selective intracellular targeting of DOR was observed in nerve growth factor (NGF)-differentiated PC12 neurosecretory cells but not in PC12 cells cultured in the absence of NGF, where both DOR and MOR were localized in the plasma membrane. Surprisingly, NGF initiated intracellular targeting of DOR in PC12 cells acutely, within 60 min after initial activation of TrkA. The NGF-induced intracellular pool of DOR originated from a late stage of the biosynthetic pathway after exit from the endoplasmic reticulum and processing of N-linked glycans in the Golgi, resulting in the accumulation in cells of a biochemically mature "reserve" pool of intracellular DOR that exhibited depolarization-dependent insertion into the plasma membrane. The C-terminal cytoplasmic tail of DOR contains a signal determining the specificity of NGF-regulated intracellular targeting. These results indicate that cloned opioid receptors are differentially targeted when expressed heterologously in neurosecretory cells, establish a model system that facilitates mechanistic study of this process, and suggest a novel function of neurotrophins in modulating the anterograde membrane trafficking of opioid receptors.
Kim
Kyung-Ah
KA
Department of Psychiatry, University of California, San Francisco School of Medicine, San Francisco, California 94143-0984, USA.
von Zastrow
Mark
M
D000818:Animals; D002451:Cell Compartmentation / Q000187:drug effects; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D002478:Cells, Cultured; D005089:Exocytosis; D006801:Humans; D007425:Intracellular Membranes / Q000378:metabolism; D051379:Mice; D020932:Nerve Growth Factor / Q000494:pharmacology / Q000502:physiology; D009414:Nerve Growth Factors / Q000494:pharmacology / Q000502:physiology*; D009473:Neuronal Plasticity / Q000502:physiology; D009474:Neurons / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D009490:Neurosecretory Systems / Q000166:cytology / Q000378:metabolism*; D016716:PC12 Cells; D017434:Protein Structure, Tertiary / Q000502:physiology; D021381:Protein Transport / Q000187:drug effects / Q000502:physiology; D051381:Rats; D017465:Receptors, Opioid, delta / Q000235:genetics / Q000378:metabolism*; D017450:Receptors, Opioid, mu / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction / Q000502:physiology; D014162:Transfection; D021601:trans-Golgi Network / Q000187:drug effects / Q000378:metabolism
D009414:Nerve Growth Factors; D017465:Receptors, Opioid, delta; D017450:Receptors, Opioid, mu; D011994:Recombinant Proteins; D020932:Nerve Growth Factor