Regulation of the membrane estrogen receptor-α

Role of cell density, serum, cell passage number, and estradiol

Celeste Finnerty, Nataliya Bulayeva, David B. Brown, Bahiru Gametchu, Cheryl S. Watson

Research output: Contribution to journalArticle

37 Citations (Scopus)

Abstract

We used modified immunocytochemical conditions to quantify a membrane form of estrogen receptor-α (mERα) in a rat pituitary tumor cell line, GH3/B6/F10. We studied the regulation of mERα vs. levels of intracellular ERα (iERα) using our 96-well plate immunoassay. The anti-ERα antibody C542 was used to label the ERα (via conjugated alkaline phosphatase) in fixed permeabilized (for iERα) vs. nonpermeabilized cells (for mERα). Expression of mERα was highest at low cell densities (<1000 cells/well) and decreased significantly at densities where cellular processes touched, whereas the more abundant iERα increased with increasing cell density over the same range. Serum starvation for 48 h caused increases in mERα, whereas iERα levels showed no significant changes. A large decline in mERα and iERα levels with cell passage number was observed. Minutes after nM 17β-estradiol (E2) treatment, a portion of the cells rounded up and detached from the culture plate, whereas nM cholesterol had no such effect. Although E2 treatment did not change mERα levels, the antigen was reorganized from a fine particulate to aggregation into asymmetric large granules of staining. That common culturing conditions favor down-regulation of mERα may explain the relatively few reports of this protein in other experimental systems.

Original languageEnglish (US)
Pages (from-to)1917-1927
Number of pages11
JournalFASEB Journal
Volume16
Issue number14
DOIs
StatePublished - Dec 2002

Fingerprint

Estrogen Receptors
estradiol
Estradiol
Cell Count
Membranes
Serum
cells
estrogen receptors
Pituitary Neoplasms
Starvation
Tumor Cell Line
immunoassays
Immunoassay
starvation
Alkaline Phosphatase
alkaline phosphatase
cell membranes
Rats
Labels
Tumors

Keywords

  • Estrogens
  • Nongenomic effects of steroids
  • Nuclear receptors
  • Rapid

ASJC Scopus subject areas

  • Agricultural and Biological Sciences (miscellaneous)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Biochemistry
  • Cell Biology

Cite this

Regulation of the membrane estrogen receptor-α : Role of cell density, serum, cell passage number, and estradiol. / Finnerty, Celeste; Bulayeva, Nataliya; Brown, David B.; Gametchu, Bahiru; Watson, Cheryl S.

In: FASEB Journal, Vol. 16, No. 14, 12.2002, p. 1917-1927.

Research output: Contribution to journalArticle

Finnerty, Celeste ; Bulayeva, Nataliya ; Brown, David B. ; Gametchu, Bahiru ; Watson, Cheryl S. / Regulation of the membrane estrogen receptor-α : Role of cell density, serum, cell passage number, and estradiol. In: FASEB Journal. 2002 ; Vol. 16, No. 14. pp. 1917-1927.
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