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The Journal of Neuroscience, January 1, 1999, 19(1):56-63
Requirement of Receptor Internalization for Opioid Stimulation of Mitogen-Activated Protein Kinase: Biochemical and Immunofluorescence Confocal Microscopic Evidence
Elena G. Ignatova, Mariana M. Belcheva, Laura M. Bohn, Mark C. Neuman, and Carmine J. Coscia E. A. Doisy Department of Biochemistry and Molecular Biology, St. Louis University School of Medicine, St. Louis, Missouri 63104
Previously, we implicated the opioid receptor (OR), G
subunits, and Ras in the opioid activation of extracellular signal-regulated protein kinase (ERK), a member of the mitogen-activated protein (MAP) kinase family involved in mitogenic signaling. We now report that OR endocytosis also plays a role in the opioid stimulation of ERK activity. COS-7 and HEK-293 cells were cotransfected with the cDNA of
-, µ-, or
-OR, dynamin wild-type (DWT), or the dominant suppressor mutant dynamin K44A, which blocks receptor endocytosis. The activation of ERK by opioid agonists in the presence of DWT was detected. In contrast, parallel ectopic coexpression of the K44A mutant with OR, followed by agonist treatment, resulted in a time-dependent attenuation of ERK activation. Immunofluorescence confocal microscopy of
Key words: opioid receptor; opioids; MAP kinase; dynamin; ERK; immunofluorescence confocal microscopy
Copyright © 1999 Society for Neuroscience 0270-6474/99/19156-08$05.00/0
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