Kelton C A, Cheng S V, Nugent N P, Schweickhardt R L, Rosenthal J L, Overton S A, Wands G D, Kuzeja J B, Luchette C A, Chappel S C
Ares Advanced Technology, Inc., Randolph, MA 02368.
Mol Cell Endocrinol. 1992 Nov;89(1-2):141-51. doi: 10.1016/0303-7207(92)90220-z.
Follicle stimulating hormone (FSH) receptor clones were isolated from a human testis cDNA library. Characterization of the cDNA clones showed that the DNA and predicted amino acid sequences of the long open reading frame differed from a previously published human ovarian FSH receptor sequence (Minegish et al. (1991) Biochem. Biophys. Res. Commun. 175, 1125-1130) by seven nucleotides and five amino acids. A human FSH receptor splice variant was also identified and characterized. A full-length human FSH receptor cDNA was engineered for expression in COS-7, CHO, and Y-1 cells. In transient transfections of COS-7 cells and stable transfections of Y-1 cells, efficient FSH receptor mRNA accumulation and isolation of FSH-responsive cell lines occurred only when an intron was included in the 5' untranslated region of the FSH receptor transcription unit. Y-1 cells stably transfected with the FSH receptor responded to FSH treatment by rounding up and by synthesizing increased amounts of progesterone. Stably transfected CHO cell lines, which responded to FSH by synthesizing increased amounts of cAMP, were isolated irrespective of the presence of the heterologous intron. The FSH-responsive CHO and Y-1 cell lines may be suitable for the development of better in vitro FSH bioassays. These cells also constitute a convenient source of human FSH receptor protein for use in radioreceptor assays and in studies of receptor-ligand interactions.
从人睾丸cDNA文库中分离出促卵泡激素（FSH）受体克隆。对cDNA克隆的表征显示，长开放阅读框的DNA和预测的氨基酸序列与先前发表的人卵巢FSH受体序列（Minegish等人，（1991年）《生物化学与生物物理学研究通讯》175，1125 - 1130）有七个核苷酸和五个氨基酸的差异。还鉴定并表征了一种人FSH受体剪接变体。构建了全长人FSH受体cDNA以在COS - 7、CHO和Y - 1细胞中表达。在COS - 7细胞的瞬时转染和Y - 1细胞的稳定转染中，仅当FSH受体转录单位的5'非翻译区包含一个内含子时，才会出现有效的FSH受体mRNA积累并分离出FSH反应性细胞系。用FSH受体稳定转染的Y - 1细胞对FSH处理的反应是细胞变圆并合成增加量的孕酮。无论是否存在异源内含子，都分离出了通过合成增加量的cAMP对FSH有反应的稳定转染的CHO细胞系。FSH反应性CHO和Y - 1细胞系可能适用于开发更好的体外FSH生物测定法。这些细胞也构成了用于放射受体测定和受体 - 配体相互作用研究的人FSH受体蛋白的便利来源。
