Expression of the neuropeptide Y Y-1 receptor in human embryonic kidney 293 cells: Ligand binding characteristics, in situ hybridization and receptor autoradiography

被引:13
|
作者
Tong, Y
Dumont, Y
Shen, SH
Herzog, H
Shine, J
Quirion, R
机构
[1] MCGILL UNIV,FAC MED,DOUGLAS HOSP,RES CTR,VERDUN,PQ H4H 1R3,CANADA
[2] MCGILL UNIV,FAC MED,DEPT PSYCHIAT,VERDUN,PQ H4H 1R3,CANADA
[3] NATL RES COUNCIL CANADA,BIOTECHNOL RES INST,MONTREAL,PQ H4P 2R2,CANADA
[4] ST VINCENTS HOSP,GARVAN INST MED RES,DARLINGHURST,NSW 2010,AUSTRALIA
来源
MOLECULAR BRAIN RESEARCH | 1995年 / 34卷 / 02期
基金
英国医学研究理事会;
关键词
neuropeptide Y Y-1 receptor; transfected cells; in situ hybridization; autoradiography;
D O I
10.1016/0169-328X(95)00176-S
中图分类号
Q189 [神经科学];
学科分类号
071006 ;
摘要
The human neuropeptide Y Y-1 receptor cDNA was transfected into human embryonic kidney 293 (HEK 293) cells and used to determine the selectivity of newly developed Y-1 and Y-2 radioligands in a model which expresses a single NPY receptor subtype. The Y-1 receptor probe, [I-125][Leu(31),Pro(34)]PYY, binds with high afinity (K-D of 0.4-0.6 nM) to Y-1-transfected HEK 293 cells whereas the Y-2 radioligand, [I-125]PYY3-36 failed to demonstrate any signficant labelling. Only non-selective (PYY) or selective Y-1 receptor agonists behaved as potent competitors for [I-125][Leu(31),Pro(34)]PYY binding in transfected cells. Additionally, the efficacy of the transfection method used was evaluated at both the transcriptional and translational levels. In situ hybridization revealed the heterogeneous distribution of the NPY Y, receptor mRNA expressed in transfected HEK 293 cells. Similarly, the levels of NPY Y-1 binding sites per transfected cell varied as shown using [I-125][Leu(31),pro(34)]PYY receptor autoradiography. Taken together, these results demonstrate that HEK 293 cells transfected with the NPY Y-1 receptor cDNA expressed both the related receptor mRNA and protein albeit at different levels depending upon each transfected cell. Additionally, these data further establish the selectivity of the newly developed Y-1 and Y-2 radioligands.
引用
收藏
页码:303 / 308
页数:6
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