Abbreviations used : β2-AR, β2-adrenergic receptor ; CP 55,940, (1α, 2β)-(R)-5-(1,1-dimethylheptyl)-2-[5-hydroxy-2-(3-hydroxy-2-(3-hydroxypropyl)cyclohexyl]phenol ; CTX, cholera toxin ; GPCR, G protein-coupled receptor ; GRK, G protein receptor kinase ; GRP, G protein receptor phosphatase ; HU 210, 11-hydroxy-Δ8-dimethylheptyltetrahydrocannabinol ; PB, phosphate buffer ; PBS, phosphate-buffered saline ; PTX, pertussis toxin ; SR 141716A, N-(piperidin-1-yl)-5-(4-chlorophenyl)-1-(2,4-dichlorophenyl)-4-methyl-1H-pyrazole-3-carboxamide hydrochloride ; Δ9-THC, Δ9-tetrahydrocannabinol ; WIN 55,212-2, R-(+)-(2,3-dihydro-5-methyl-3-[(4-moropholinyl)methyl]pyrol[1,2,3-de]-1,4-benzoxazin-6-yl)(1-naphthalenyl)methanone monomethanesulfonate.
Internalization and Recycling of the CB1 Cannabinoid Receptor
Article first published online: 18 JAN 2002
Journal of Neurochemistry
Volume 73, Issue 2, pages 493–501, August 1999
How to Cite
Hsieh, C., Brown, S., Derleth, C. and Mackie, K. (1999), Internalization and Recycling of the CB1 Cannabinoid Receptor. Journal of Neurochemistry, 73: 493–501. doi: 10.1046/j.1471-4159.1999.0730493.x
- Issue published online: 18 JAN 2002
- Article first published online: 18 JAN 2002
- CB1 cannabinoid receptor;
- Cannabimimetic drugs;
- Receptor regulation.
Abstract : Tolerance develops rapidly to cannabis, cannabinoids, and related drugs acting at the CB1 cannabinoid receptor. However, little is known about what happens to the receptor as tolerance is developing. In this study, we have found that CB1 receptors are rapidly internalized following agonist binding and receptor activation. Efficacious cannabinoid agonists (WIN 55,212-2, CP 55,940, and HU 210) caused rapid internalization. Methanandamide (an analogue of an endogenous cannabinoid, anandamide) was less effective, causing internalization only at high concentration, whereas Δ9-tetrahydrocannabinol caused little internalization, even at 3 μM. CB1 internalized via clathrin-coated pits as sequestration was inhibited by hypertonic sucrose. Internalization did not require activated G protein αi, αo, or αs subunits. A region of the extreme carboxy terminus of the receptor was necessary for internalization, as a mutant CB1 receptor lacking the last 14 residues did not internalize, whereas a mutant lacking the last 10 residues did. Steps involved in the recycling of sequestered receptor were also investigated. Recovery of CB1 to the cell surface after short (20 min) but not long (90 min) agonist treatment was independent of new protein synthesis. Recycling also required endosomal acidification and dephosphorylation. These results show that CB1 receptor trafficking is dynamically regulated by cannabimimetic drugs.