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Vol. 60, Issue 6, 1195-1200, December 2001
2C-Adrenoceptors
from Golgi to Plasma Membrane in Transfected Human Embryonic Kidney 293 Cells
Dorothy M. Davis Heart and Lung Research Institute, The Ohio State
University, Columbus, Ohio
Cold-induced vasoconstriction in cutaneous blood vessels is mediated by
increased constrictor activity of vascular
2-adrenoceptors (2-ARs). In mouse
cutaneous arteries, 2-AR constriction at 37°C is
mediated by 2A-ARs, whereas after cold exposure
(28°C), 2C-ARs are no longer silent and mediate the
remarkable cold-induced augmentation of 2-AR
responsiveness. The goals of the present study were to develop a cell
model of cutaneous thermoregulation and to determine the mechanisms
underlying the thermosensitivity of 2C-ARs. Human embryonic kidney 293 cells were transiently transfected with the mouse
2A- or 2C-AR. In cells expressing
2A-ARs, UK-14,304
(5-bromo-N-(4,5-dihydro-1H-imidazol-2-yl)-6-quinoxalinamine), an 2-AR agonist, inhibited (10 pM) and stimulated (1-10
nM) the accumulation of cAMP evoked by forskolin. Similar responses
were obtained at 37°C and 28°C. In contrast, in cells expressing
2C-ARs, UK-14,304 did not affect forskolin-stimulated
cAMP accumulation at 37°C but did cause a concentration-dependent
inhibitory effect at 28°C. Subcellular fractionation revealed that at
37°C 2C-ARs were localized predominantly to Golgi
compartments, whereas 2A-ARs localized predominantly to
the plasma membrane. After cooling (28°C), 2C-ARs
relocated from Golgi compartments to the plasma membrane, whereas the
2A-AR remained at the plasma membrane. Immunofluorescence microscopy confirmed that, at 37°C,
2A-ARs were localized to the cell surface, whereas
2C-ARs colocalized with a trans-Golgi
marker. Cooling did not affect localization of 2A-ARs,
but shifted 2C-ARs to the cell surface. Moderate cooling, therefore, caused a selective redistribution of
2C-ARs from the Golgi compartments to the cell surface,
allowing the rescue of the 2C-adrenergic functional
response. This mechanism may explain the role of 2-ARs
in thermoregulation of the cutaneous circulation.
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