title: Desensitization and resensitization of lutropin receptors expressed in transfected Y-1 adrenal cells.
abstract: Stimulation of gonadal cells by lutropins such as human chorionic gonadotropin (hCG) is often transient and followed by down-regulation and/or desensitization of lutropin receptors (LHR). Here we describe desensitization/resensitization of LHR in Y-1 adrenal cell lines (termed Y-1L) expressing a rat cDNA lacking most 5' and 3' LHR untranslated regions under the control of a metallothionein promoter. Using a simple morphological assay in which stimulated cells are round and unstimulated cells are flat, we identified clones that rounded and remained round and others that became insensitive to lutropin stimulation and reverted to their flat appearance within 2-4 h. Flattened cells were insensitive to further hormonal stimulation but rounded after treatments with cholera toxin, forskolin, or cyclic AMP, showing that loss of responsiveness was associated with an early step in signal transduction, not loss of rounding potential. Removing the lutropin stimulus for at least 90-120 min reversed hormone insensitivity, even in the presence of the protein synthesis inhibitor puromycin. The number of surface bound receptors did not change during a cycle of rounding/flattening and hCG bound to rounded or flattened cells was replaced equally by radioiodinated hCG during incubations at 4 degrees C. Thus, desensitization/resensitization of LHR in Y-1L cells occurred in the absence of new receptor synthesis, receptor degradation, or receptor recycling. These observations suggest that LHR desensitization/resensitization in Y-1L cells was closely coupled to receptor occupancy and that this cell line may be useful for identifying factors that modulate the activities of occupied receptors.
authors.affiliation: Department of OBGYN, Robert Wood Johnson Medical School, 675 Hoes Lane, Piscataway, New Jersey 08854, USA.
authors.forename: G A
authors.initials: GA
authors.lastname: Ulaner
authors.forename: J
authors.initials: J
authors.lastname: Chuang
authors.forename: W
authors.initials: W
authors.lastname: Lin
authors.forename: D
authors.initials: D
authors.lastname: Woodbury
authors.forename: R V
authors.initials: RV
authors.lastname: Myers
authors.forename: W R
authors.initials: WR
authors.lastname: Moyle
mesh_terms: D000311:Adrenal Glands / Q000737:chemistry / Q000166:cytology / Q000187:drug effects*; D000818:Animals; D002460:Cell Line; D048429:Cell Size / Q000187:drug effects; D006063:Chorionic Gonadotropin / Q000494:pharmacology; D000242:Cyclic AMP / Q000378:metabolism; D015536:Down-Regulation; D007986:Luteinizing Hormone / Q000494:pharmacology; D008855:Microscopy, Electron, Scanning; D051381:Rats; D011974:Receptors, LH / Q000378:metabolism*; D014162:Transfection
chemical_list: D006063:Chorionic Gonadotropin; D011974:Receptors, LH; D007986:Luteinizing Hormone; D000242:Cyclic AMP