H. Boudin et al., Somatostatin-induced regulation of SST2A receptor expression and cell surface availability in central neurons: Role of receptor internalization, J NEUROSC, 20(16), 2000, pp. 5932-5939
To investigate the effects of somatostatin (somatotropin release-inhibiting
factor, SRIF) on the regulation of SST2A receptors in mammalian brain, we
examined how blockade of SRIF release or stimulation by the SRIF analog [D-
Trp(8)]-SRIF would affect the expression and cell surface availability of S
ST2A receptors in rat brain slices. First, we measured the intensity of SST
2A immunoreactivity, using quantitative light microscopic immunocytochemist
ry, and levels of SST2A mRNA, using semiquantitative RT-PCR, under conditio
ns of acute SRIF release blockade. Incubation of slices from the claustrum
or basolateral amygdala, two regions previously shown to contain high conce
ntrations of SST2A receptors, in Ca2+-free Ringer's for 40 min induced a de
crease in the intensity of SST2A receptor immunoreactivity and concentratio
n of SST2A mRNA as compared with control values obtained in Ca2+-supplement
ed Ringer's. These effects were counteracted in a dose-dependent manner by
the addition of 10-100 nM [D-Trp(8)]-SRIF to the Ca2+-free medium. Furtherm
ore, both of these effects were abolished in the presence of the endocytosi
s inhibitors phenylarsine oxide or hyperosmolar sucrose, suggesting that th
ey were dependent on receptor internalization. Electron microscopic immunog
old labeling confirmed the existence of an agonist-induced internalization
of SST2A receptors in central neurons. At a high (10 mu M), but not at a lo
w (10 nM), concentration of agonist this internalization resulted in a sign
ificant decrease in cell surface receptor density, irrespective of the pres
ence of Ca2+ in the medium. Taken together, these results suggest that liga
nd-induced endocytosis is responsible for rapid transcriptional (increase i
n SST2A expression) and trafficking (loss of cell surface receptors) events
involved in the control of the somatostatinergic signal.