Mt. Teh et D. Sugden, The putative melatonin receptor antagonist GR128107 is a partial agonist on Xenopus laevis melanophores, BR J PHARM, 126(5), 1999, pp. 1237-1245
1 GR128107 (3-(1-acetyl-3-methyl-piperidine)-5-methoxyindole) has previousl
y been reported to be a competitive melatonin receptor antagonist in blocki
ng melatonin inhibition of [H-3]-dopamine release from rabbit retina, a res
ponse mediated by the MT2 receptor subtype.
2 GR128107, like melatonin, induced a rapid (maximum response in 60-90 min)
pigment aggregation in a clonal line of Xenopus laevis melanophores. GR128
107 behaved as a partial agonist (pEC(50) 8.58+/-0.03, n=3) with an E-max o
f 0.83 (relative to melatonin, pEC(50) 10.09+/-0.03, n=3).
3 The concentration-response curve for pigment granule aggregation to both
melatonin and GR128107 was displaced in a parallel, rightward manner by mel
atonin receptor antagonists with very similar potencies; estimated pK(B) RJ
252 (against melatonin 4.60/against GR128107 4.54) < GR135533 (6.40/6.14) <
Luzindole (6.45/6.49) < S20929 (6.58/6.65) < 4-P-PDOT (6.73/6.85).
4 Bath melatonin- and GR128107-induced pigment granule aggregation was prev
ented by pretreatment of melanophores with pertussis toxin (10-1000 ng ml(-
1)).
5 Prolonged pre-treatment of melanophores with melatonin desensitized the p
igment aggregation response to GR128107. In desensitized cells, the maximal
aggregation produced by GR128107 was only 0.27+/-0.01 (n=4) and the pEC(50
) was reduced (vehicle 8.57+/-0.12; melatonin pre-treated 7.84+/-0.09, n=4)
. The maximal response to melatonin in desensitized melanophores was unchan
ged but the pEC(50) was reduced (vehicle 10.49+/-0.03; melatonin pre-treate
d 9.83+/-0.04, n=4).
6 These results demonstrate that GR128107 induces pigment granule aggregati
on in Xenopus melanophores by activating a cell membrane melatonin receptor
coupled via a pertussis toxin-sensitive G-protein.
7 The partial agonist activity of GR128107 in melanophores may be apparent
because of the very high density of melatonin receptors in these cells (B-m
ax 1223 fmol mg protein(-1)) compared to the low density of sites in rabbit
retina (B-max 3.1 fmol mg protein(-1)). This suggestion is supported by th
e finding that GR128107, like melatonin, acted as a full agonist and inhibi
ted forskolin-stimulation of cyclic AMP accumulation in NIH-3T3 cells expre
ssing a high density of human mt(1) or MT2 receptors.