Visualization of unoccupied and occupied gonadotropin-releasing hormone receptors in living cells

Xinwei Lin, Anda Cornea, Jo Ann Janovick, P. Michael Conn

    Research output: Contribution to journalArticle

    16 Scopus citations

    Abstract

    Three chimeras of the rat GnRH receptor (rGnRHR) and an enhanced green fluorescent protein (GFP) were assessed to examine their suitability as probes of the receptor in transfected GH3 cells. Direct fusion of GFP to the N or C terminus of the rGnRHR abolished the receptor ligand binding affinity and the chimeric receptors were intracellularly localized. In contrast, rGnRHR-Ctail-GFP, a fusion of the N-terminus of the GFP to the C-terminus of the rGnRHR with the intracellular C-terminal tail of the catfish GnRHR as an intermediate spacer, was functional in terms of plasma membrane localization, ligand binding ability, receptor-mediated signal transduction and pattern of homologous down-regulation. The functional chimera of GnRHR and GFP provided a useful model for observation of GnRHR distribution and agonist-stimulated trafficking in living cells.

    Original languageEnglish (US)
    Pages (from-to)27-37
    Number of pages11
    JournalMolecular and Cellular Endocrinology
    Volume146
    Issue number1-2
    DOIs
    StatePublished - Nov 25 1998

    Keywords

    • Confocal microscopy
    • GnRH recepfor
    • Green fluorescent protein

    ASJC Scopus subject areas

    • Biochemistry
    • Molecular Biology
    • Endocrinology

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