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.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Berthois, Y., Pourreau-Schneider, N., Gandilhon, P., Mittre, H., Tubiana, N. & Martin, P.M. (1986).J. Steroid Biochem.,25, 963–972.
Bression, D., Michard, M., Le Dafniet, M., Pagesy, P. & Peillon, F. (1986).Endocr.,119, 1048–1051.
Cole, S.R., Ashman, L.K. & Ely, P.L. (1987).Molecular Immunology,24, 699–705.
Dannies, P.S. (1985). In:Biochemical Actions of Hormones. Litwack, G. (ed.) pp. 289–310. Academic Press, Inc: Orlando.
Denechaud, M., Israel, J., Mishal, Z. & Vincent, J. (1987).Life Sciences,40, 2377–2384.
Dufy, B., Partouche, C., Poulain, D., Dufy-Barbe, L. & Vincent, J.D. (1976).Neuroenclocrinology,22, 38–47.
Dufy, B., Vincent, J.-D., Fleury, H., Pasquier, P.D., Gourdji, D. & Vidal, A.T. (1979).Science,204, 509–511.
Gametchu, B. (1987).Science,236, 456–461.
Gametchu, B., Watson, C., Shih, C. & Dashew, B. (1991a).Steroids,56, 411–419.
Gametchu, B., Watson, C.S. & Pasko, D. (1991b).Steroids,56, 402–410.
Gametchu, B., Watson, C.S. & Wu, S. (1993).FASEB J.,7, 1283–1293.
Gametchu, B., Chen, F. & Watson, C.S. (1995). In:Glucocorticoid Receptor Structure and Leukemic Cell Responses. Gametchu, B. (ed.) pp. 75–103. R.G. Landes Co: Austin.
Gametchu, B. & Watson, C.S. (1995). In:Glucocorticoid Receptor Structure and Leukemic Cell Responses. Gametchu, B. (ed.) pp. 163–176. R.G. Landes Company: Austin.
Gautvik, K.M. & Fossum, S. (1976).Biochem. J.,158, 119–125.
Hayashi, I. & Sato, G.H. (1976).Nature,259, 132–134.
Hoffman, M. (1991).Science,254, 650–651.
Ibarrolla, I., Alejandro, A., Marino, A., Sancho, M.I. & Trueba, M. (1992).J. Membr. Biol,125, 185–191.
Kelly, M.J., Moss, R.L. & Dudley, C.A. (1977).Exp. Brain Res.,30, 53–64.
Klein, P., Kanehisa, M. & DeLisi, C. (1985).Biochim. Biophys Acta,815, 468–476.
Labarca, C. & Paigen, K. (1980).Anal. Biochem.,102, 344–352.
Lieberherr, M., Grosse, B., Kachkache, M. & Balsan, S. (1993).J. Bone Min. Res.,8, 1365–1376.
Morley, P., Whitfield, J.F., Vanderhyden, B.C., Tsang, B.K. & Schwartz, J. (1992).Endocr.,131, 1305–1312.
Nabekura, J., Oomura, Y. Minami, T., Mizuno, Y. & Fukuda, A. (1986).Science,233, 226–228.
Nenci, I., Fabris, G., Marchetti, E. & Marzola, A. (1980). In:Perspectives in Steroid Receptor Research. Bresciani, F. (ed.) pp. 61–72. Raven Press: New York.
Neter, J. & Wasserman. W. (1974). In: Applied Linear Statistical Models. Richard E. Irwin, Inc. Homewood, IL.
Pappas, T.C., Gametchu. B., Yannariello-Brown, J., Collins, T.J. & Watson, C.S. (1994).Endocrine,2, 813–822.
Pappas, T.C., Gametchu. B. & Watson, C.S. (1995).FASEB J.,9, 404–410.
Pietras, R.J. & Szego, C.M. (1977).Nature,265, 69–72.
Pietras, R.J. & Szego, C.M. (1979a).J. Steroid. Biochem.,11, 1471–1483.
Pietras, R.J. & Szego, C.M. (1979b).J. Cell. Physiol.,98, 145–160.
Pietras, R.J. & Szego, C.M. (1980).Biochem. J.,191, 743–760.
Pourreau-Schneider, N., Berthois, Y., Gandihon, P., Cau, P., Martin, P.M. & Passerel, M. (1986).Mol. Cell Endocrinol.,48, 77–88.
Scheffé, H. (1959). In: The Analysis of Variance. John Wiley & Sons: New York.
Towle, A.C. & Sze, P.Y. (1983).J. Steroid Biochem.,18, 135–143.
Turksen, K. & Aubin, J.E. (1994).J. Cell Biol.,114, 373–384.
Warrington, A.E., Barbarese, E. & Pfeiffer, S.E. (1992).Dev. Neurosci.,14, 93–97.
Weigel, P. (1980).J. Biol. Chem.,255, 6111–6120.
Weigel, P.H. & Oka, J.A. (1983).J. Biol. Chem.,258, 5089–5094.
Weissman, A. (1991). In:Current Protocols in Immunology. Coligan, J.E., Kruisbeek, A.M., Margulies, D.H., Shevach, E.M. & Strober, W. (eds.) pp. 8.1.1–8.1.9. John Wiley and Sons: New York.
Welshons, W.V. & Judy B.M. (1995).Endocrine,3, 1–4.
Wong, M. & Moss, R.L. (1992).J. Neurosci.,12, 3217–3225.
Zyzek, E., Dufy-Barbe, L., Dufy, B. & Vincent, J.D. (1981).Biochem. Biophys. Res. Commun.,102, 1151–1157.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Pappas, T.C., Gametchu, B. & Watson, C.S. Membrane estrogen receptor-enriched GH3/B6 cells have an enhanced non-genomic response to estrogen. Endocr 3, 743–749 (1995). https://doi.org/10.1007/BF03000207
Issue Date:
DOI: https://doi.org/10.1007/BF03000207