The cloning of the human follicle stimulating hormone receptor and its expression in COS-7, CHO, and Y-1 cells

https://doi.org/10.1016/0303-7207(92)90220-ZGet rights and content

Abstract

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 recéptor-ligand interactions.

References (44)

  • P. Chomczynski et al.

    Anal. Biochem.

    (1987)
  • A.P. Feinberg et al.

    Anal. Biochem.

    (1983)
  • F. Libert et al.

    Biochem. Biophys. Res. Commun.

    (1989)
  • T. Minegish et al.

    Biochem. Biophys. Res. Commun.

    (1990)
  • T. Minegish et al.

    Biochem. Biophys. Res. Commun.

    (1991)
  • J. Nathans et al.

    Cell

    (1983)
  • C.H. Tsai-Morris et al.

    J. Biol. Chem.

    (1990)
  • Y.-B. Xie et al.

    J. Biol. Chem.

    (1990)
  • T. Braun et al.

    EMBO J.

    (1991)
  • A.R. Buchman et al.

    Mol. Cell. Biol.

    (1988)
  • J. Callis et al.

    Genes Dev.

    (1987)
  • G.M. Church et al.
  • S. Cotecchia et al.
  • R.A.F. Dixon et al.

    Nature

    (1986)
  • J.C. Fiddes et al.

    J. Mol. Appl. Genet

    (1981)
  • J.B. Field
  • A.L. Frazier et al.

    Mol. Endocrinol.

    (1990)
  • T. Frielle et al.
  • I.B. Fritz et al.

    Can. J. Biochem.

    (1978)
  • I.B. Fritz et al.

    Natl. Cancer Inst. Monogr.

    (1978)
  • L.H. Graff et al.

    Mol. Cell. Biol.

    (1982)
  • D.H. Hamer et al.

    J. Mol. Appl. Genet.

    (1982)
  • Cited by (150)

    • Gonadotropin Hormones and Their Receptors

      2018, Yen & Jaffe's Reproductive Endocrinology: Physiology, Pathophysiology, and Clinical Management: Eighth Edition
    • Towards a non-animal risk assessment for anti-androgenic effects in humans

      2015, Environment International
      Citation Excerpt :

      These factors explain why there are no widely reported standardized assays for characterizing the effects of chemicals on the FSHR. However, recombinant human FSHR have been successfully transfected into both rodent (Kelton et al., 1992; Christin-Maitre and Bouchard, 1996) and human (HEK293) cells (Karakaya et al., 2014). Both LHR and FSHR are expressed on some human cancer cell lines such as OCC1, and this cell line proliferates in response to stimulation by FSH or hCG (Parrott et al., 2001), thus providing a potential assay of in vitro physiological response for agonism or antagonism at these receptors.

    • Structural biology of glycoprotein hormones and their receptors: Insights to signaling

      2014, Molecular and Cellular Endocrinology
      Citation Excerpt :

      Indeed, thyrostimulin, which lacks a putative ‘seatbelt’ loop, is not as stable as the other GPHs (Okajima et al., 2008). The advances in cloning and expression of glycoprotein hormone receptors (GPHRs) since 1989 (Kelton et al., 1992; McFarland et al., 1989; Parmentier et al., 1989; Sprengel et al., 1990) initiated a period of rapid progress in understanding and ultimately providing reagents which would lead to the structural evidence for the mode of interaction of GPH ligands with their receptors. An imperfect motif of leucine-rich repeats (LRRs) in the sequence of the ectodomain of LHR identified by McFarland and coworkers led to the proposal that the ectodomain contains 14 LRRs (McFarland et al., 1989).

    View all citing articles on Scopus
    1

    Current address: Diacrin, Bldg. 96, 13th Street, Charlestown, MA 02109, USA.

    View full text