TY - JOUR
T1 - Colchicine
T2 - A novel positive allosteric modulator of the human 5-Hydroxytryptamine 3A receptor
AU - De Oliveira-Pierce, A. N.
AU - Zhang, R.
AU - Machu, T. K.
PY - 2009/5
Y1 - 2009/5
N2 - The actions of colchicine were examined with the two-electrode voltage-clamp technique and radioligand binding assays in mouse and human 5-hydroxytryptamine 3A receptors (5-HT 3ARs) expressed in Xenopus laevis oocytes. Colchicine inhibited 5-hydroxytryptamine (5-HT)-evoked currents in oocytes expressing mouse 5-HT 3ARs, with an IC 50 of 59.5 ± 3 μM. In contrast to the mouse receptor, coapplication of colchicine with 5-HT (<1 μM) strongly enhanced 5-HT-evoked currents in oocytes expressing human 5-HT 3ARs. Colchicine applied alone did not induce a detectable current. In the presence of 0.5 μM 5-HT, the potentiation was concentration-dependent and reached the maximum (∼100%) when 750 μM colchicine was applied. However, colchicine-dependent inhibition can be observed at 5-HT concentrations > 1 μM. In oocyte membranes expressing mouse or human receptors, binding studies with colchicine (25 nM-1 mM) revealed no displacement of 1-methyl-N-((1R,3r,5S)-9-methyl-9 azabicyclo [3.3.1]nonan-3yl)-1H-indazole-3 carboxamide ([ 3H]BRL- 43694), suggesting that actions of colchicine do not occur at the ligand binding domain. Functional effects of colchicine on both receptors occurred in the absence of preincubation and after cold temperature incubation, suggesting that the microtubule-depolymerizing effects of colchicine play no role in modulation of receptor function. Studies with interspecies chimeric receptors demonstrated that the distal one third of the N terminus is responsible for the bidirectional modulation by colchicine. Collectively, these results suggest that colchicine modulates receptor function through loops C and/or F through a gating mechanism.
AB - The actions of colchicine were examined with the two-electrode voltage-clamp technique and radioligand binding assays in mouse and human 5-hydroxytryptamine 3A receptors (5-HT 3ARs) expressed in Xenopus laevis oocytes. Colchicine inhibited 5-hydroxytryptamine (5-HT)-evoked currents in oocytes expressing mouse 5-HT 3ARs, with an IC 50 of 59.5 ± 3 μM. In contrast to the mouse receptor, coapplication of colchicine with 5-HT (<1 μM) strongly enhanced 5-HT-evoked currents in oocytes expressing human 5-HT 3ARs. Colchicine applied alone did not induce a detectable current. In the presence of 0.5 μM 5-HT, the potentiation was concentration-dependent and reached the maximum (∼100%) when 750 μM colchicine was applied. However, colchicine-dependent inhibition can be observed at 5-HT concentrations > 1 μM. In oocyte membranes expressing mouse or human receptors, binding studies with colchicine (25 nM-1 mM) revealed no displacement of 1-methyl-N-((1R,3r,5S)-9-methyl-9 azabicyclo [3.3.1]nonan-3yl)-1H-indazole-3 carboxamide ([ 3H]BRL- 43694), suggesting that actions of colchicine do not occur at the ligand binding domain. Functional effects of colchicine on both receptors occurred in the absence of preincubation and after cold temperature incubation, suggesting that the microtubule-depolymerizing effects of colchicine play no role in modulation of receptor function. Studies with interspecies chimeric receptors demonstrated that the distal one third of the N terminus is responsible for the bidirectional modulation by colchicine. Collectively, these results suggest that colchicine modulates receptor function through loops C and/or F through a gating mechanism.
UR - http://www.scopus.com/inward/record.url?scp=65649123744&partnerID=8YFLogxK
U2 - 10.1124/jpet.108.146522
DO - 10.1124/jpet.108.146522
M3 - Article
C2 - 19188483
AN - SCOPUS:65649123744
SN - 0022-3565
VL - 329
SP - 838
EP - 847
JO - Journal of Pharmacology and Experimental Therapeutics
JF - Journal of Pharmacology and Experimental Therapeutics
IS - 2
ER -