McIlhinney R A, Molnár E, Atack J R, Whiting P J
Medical Research Council, Anatomical Neuropharmacology Unit, Oxford, U.K.
Neuroscience. 1996 Feb;70(4):989-97. doi: 10.1016/0306-4522(95)00419-x.
Cell surface expression of the NR1a subunit has been examined in mouse L cell lines permanently transfected with the complementary DNA for human NR1a or with the complementary DNAs for NR1a and NR2A. The expression of the subunits was under the control of the murine mammary tumour virus promoter and following induction of expression by dexamethazone both cell lines expressed high levels of the NR1a subunit as determined by immunofluorescence using permeabilized cells and immunoblotting of cell membranes with subunit specific antibodies. However, cell surface expression of the NR1a subunit was found only in the cells expressing both the NR1a and NR2A subunits. This was confirmed by cell surface biotinylation of the two cell lines and affinity isolation of the receptor subunits. To determine if this result was solely due to the use of a particular cell line and or the choice of expression vector, Cos-7 cells were transiently transfected with either NR1a or NR1a plus NR2A. Here too cell surface expression was only found following co-transfection of both subunits.
已在稳定转染了人NR1a互补DNA或NR1a与NR2A互补DNA的小鼠L细胞系中检测了NR1a亚基的细胞表面表达。这些亚基的表达受鼠乳腺肿瘤病毒启动子的控制,在用地塞米松诱导表达后,通过使用透化细胞的免疫荧光和用亚基特异性抗体对细胞膜进行免疫印迹分析,确定两个细胞系均表达高水平的NR1a亚基。然而,仅在同时表达NR1a和NR2A亚基的细胞中发现了NR1a亚基的细胞表面表达。通过对两个细胞系进行细胞表面生物素化以及对受体亚基进行亲和分离,证实了这一点。为了确定该结果是否仅归因于特定细胞系的使用和/或表达载体的选择,用NR1a或NR1a加NR2A瞬时转染了Cos-7细胞。同样,仅在两个亚基共转染后才发现细胞表面表达。