Membrane estrogen receptor-enriched GH3/B6 cells have an enhanced non-genomic response to estrogen

Todd C. Pappas, Bahiru Gametchu, Cheryl S. Watson

    Research output: Contribution to journalArticle

    35 Citations (Scopus)

    Abstract

    We immunoselected GH3/B6 cells for a membrane estrogen receptor (mER) using antibodies generated against the rat intracellular ER (iER). Immunocytochemistry with anti-ER antibodies revealed bright fluorescence distributed in patches over the surface of mER-enriched cells, while cells immuno-depleted for mER showed only low-level membrane immunofluorescence. Quantitation via digital image analysis confirmed that immunoenriched populations show increases in both stained cell numbers and intensity of staining. Short-term culturing with serum reversibly decreased the intensity of immunostaining in mER-enriched cells to immuno-depleted cell levels. The mER-enriched populations initially contained ∼85% immunopositive cells in defined medium, but when cultured continuously with serum gradually decline to ∼22% immunopositive cells by 10 weeks. Cells enriched for mER showed a significant increase in rapid (after 2 or 5 min) prolactin release when treated with 17β-estradiol, while mER-depleted cells lacked this response. Immunoprecipitabie membrane proteins isolated from mER-enriched cells were 60,000, 74,000 and ∼ 200,000 MW, compared to an iER size of 67,000. Therefore, the presence and level of an mER that is antigenically related to iER is correlated with the ability of GH3/B6 cells to mediate a rapid action of estrogen.

    Original languageEnglish
    Pages (from-to)743-749
    Number of pages7
    JournalEndocrine
    Volume3
    Issue number10
    DOIs
    StatePublished - Oct 1995

    Fingerprint

    Estrogen Receptors
    Estrogens
    Membranes
    Cell Membrane
    Serum
    Prolactin
    Population
    Fluorescent Antibody Technique
    Anti-Idiotypic Antibodies
    Estradiol
    Membrane Proteins
    Cell Count
    Fluorescence
    Immunohistochemistry
    Staining and Labeling
    Antibodies

    Keywords

    • estrogen receptor
    • GH/B6 cell
    • membrane
    • non-genomic
    • prolactin

    ASJC Scopus subject areas

    • Endocrinology
    • Endocrinology, Diabetes and Metabolism

    Cite this

    Membrane estrogen receptor-enriched GH3/B6 cells have an enhanced non-genomic response to estrogen. / Pappas, Todd C.; Gametchu, Bahiru; Watson, Cheryl S.

    In: Endocrine, Vol. 3, No. 10, 10.1995, p. 743-749.

    Research output: Contribution to journalArticle

    Pappas, Todd C. ; Gametchu, Bahiru ; Watson, Cheryl S. / Membrane estrogen receptor-enriched GH3/B6 cells have an enhanced non-genomic response to estrogen. In: Endocrine. 1995 ; Vol. 3, No. 10. pp. 743-749.
    @article{156c141b446041f08a3f1df0d0361f15,
    title = "Membrane estrogen receptor-enriched GH3/B6 cells have an enhanced non-genomic response to estrogen",
    abstract = "We immunoselected GH3/B6 cells for a membrane estrogen receptor (mER) using antibodies generated against the rat intracellular ER (iER). Immunocytochemistry with anti-ER antibodies revealed bright fluorescence distributed in patches over the surface of mER-enriched cells, while cells immuno-depleted for mER showed only low-level membrane immunofluorescence. Quantitation via digital image analysis confirmed that immunoenriched populations show increases in both stained cell numbers and intensity of staining. Short-term culturing with serum reversibly decreased the intensity of immunostaining in mER-enriched cells to immuno-depleted cell levels. The mER-enriched populations initially contained ∼85{\%} immunopositive cells in defined medium, but when cultured continuously with serum gradually decline to ∼22{\%} immunopositive cells by 10 weeks. Cells enriched for mER showed a significant increase in rapid (after 2 or 5 min) prolactin release when treated with 17β-estradiol, while mER-depleted cells lacked this response. Immunoprecipitabie membrane proteins isolated from mER-enriched cells were 60,000, 74,000 and ∼ 200,000 MW, compared to an iER size of 67,000. Therefore, the presence and level of an mER that is antigenically related to iER is correlated with the ability of GH3/B6 cells to mediate a rapid action of estrogen.",
    keywords = "estrogen receptor, GH/B6 cell, membrane, non-genomic, prolactin",
    author = "Pappas, {Todd C.} and Bahiru Gametchu and Watson, {Cheryl S.}",
    year = "1995",
    month = "10",
    doi = "10.1007/BF03000207",
    language = "English",
    volume = "3",
    pages = "743--749",
    journal = "Endocrine",
    issn = "0969-711X",
    publisher = "Humana Press",
    number = "10",

    }

    TY - JOUR

    T1 - Membrane estrogen receptor-enriched GH3/B6 cells have an enhanced non-genomic response to estrogen

    AU - Pappas, Todd C.

    AU - Gametchu, Bahiru

    AU - Watson, Cheryl S.

    PY - 1995/10

    Y1 - 1995/10

    N2 - We immunoselected GH3/B6 cells for a membrane estrogen receptor (mER) using antibodies generated against the rat intracellular ER (iER). Immunocytochemistry with anti-ER antibodies revealed bright fluorescence distributed in patches over the surface of mER-enriched cells, while cells immuno-depleted for mER showed only low-level membrane immunofluorescence. Quantitation via digital image analysis confirmed that immunoenriched populations show increases in both stained cell numbers and intensity of staining. Short-term culturing with serum reversibly decreased the intensity of immunostaining in mER-enriched cells to immuno-depleted cell levels. The mER-enriched populations initially contained ∼85% immunopositive cells in defined medium, but when cultured continuously with serum gradually decline to ∼22% immunopositive cells by 10 weeks. Cells enriched for mER showed a significant increase in rapid (after 2 or 5 min) prolactin release when treated with 17β-estradiol, while mER-depleted cells lacked this response. Immunoprecipitabie membrane proteins isolated from mER-enriched cells were 60,000, 74,000 and ∼ 200,000 MW, compared to an iER size of 67,000. Therefore, the presence and level of an mER that is antigenically related to iER is correlated with the ability of GH3/B6 cells to mediate a rapid action of estrogen.

    AB - We immunoselected GH3/B6 cells for a membrane estrogen receptor (mER) using antibodies generated against the rat intracellular ER (iER). Immunocytochemistry with anti-ER antibodies revealed bright fluorescence distributed in patches over the surface of mER-enriched cells, while cells immuno-depleted for mER showed only low-level membrane immunofluorescence. Quantitation via digital image analysis confirmed that immunoenriched populations show increases in both stained cell numbers and intensity of staining. Short-term culturing with serum reversibly decreased the intensity of immunostaining in mER-enriched cells to immuno-depleted cell levels. The mER-enriched populations initially contained ∼85% immunopositive cells in defined medium, but when cultured continuously with serum gradually decline to ∼22% immunopositive cells by 10 weeks. Cells enriched for mER showed a significant increase in rapid (after 2 or 5 min) prolactin release when treated with 17β-estradiol, while mER-depleted cells lacked this response. Immunoprecipitabie membrane proteins isolated from mER-enriched cells were 60,000, 74,000 and ∼ 200,000 MW, compared to an iER size of 67,000. Therefore, the presence and level of an mER that is antigenically related to iER is correlated with the ability of GH3/B6 cells to mediate a rapid action of estrogen.

    KW - estrogen receptor

    KW - GH/B6 cell

    KW - membrane

    KW - non-genomic

    KW - prolactin

    UR - http://www.scopus.com/inward/record.url?scp=0028818390&partnerID=8YFLogxK

    UR - http://www.scopus.com/inward/citedby.url?scp=0028818390&partnerID=8YFLogxK

    U2 - 10.1007/BF03000207

    DO - 10.1007/BF03000207

    M3 - Article

    VL - 3

    SP - 743

    EP - 749

    JO - Endocrine

    JF - Endocrine

    SN - 0969-711X

    IS - 10

    ER -