A nuclear binding assay to assess the biological activity of steroid receptors in isolated animal and human tissues

T. C. Spelsberg, M. L. Graham, N. J. Berg, T. Umehara, E. Riehl, C. B. Coulam, J. N. Ingle

Research output: Contribution to journalArticle

46 Citations (Scopus)

Abstract

This paper describes a nuclear binding assay (NB assay) which measures not only the presence of a steroid receptor in a tissue, but also the quantity of that receptor which is biologically active or functional, i.e. able to bind to nuclear acceptor sites. The assay involes the isolation viable cells from tissues and their incubation with an excess of radiolabeled steroid to encourage the activation and nuclear binding of all cellular receptors. The nuclei are isolated under conditions that remove unactivated (unbound) steroid-receptor complexes. This NB assay demonstrates, in both animal and human steroid target tissues, a saturable, tissue- and steroid-specific, and temperature- and time-dependent nuclear binding of radiolabeled steroids. These properties support a receptor-dependent nuclear binding of steroids. This assay is reproducible and requires relatively small amounts of tissue. The patterns of nuclear binding of the progesterone receptor, achieved with the assay in the avian oviduct model system, are shown to correlate with the nuclear binding of progesterone in vivo, the ability of the steroid to alter transcription, and the expression of a specific gene product, the protein avidin. The assay has been used to identify the existence of nonfunctional steroid receptors in endometrial and breast carcinomas. Therefore, this NB assay combined with the standard charcoal/hydroxylapatite methods of quantitating total cellular receptors should provide a means of assessing changes in the regulation of the biological activity of steroid receptors. Further, the assay should be useful to assess the ability of steroid analogs to properly activate their respective receptors for subsequent nuclear binding.

Original languageEnglish (US)
Pages (from-to)631-644
Number of pages14
JournalEndocrinology
Volume121
Issue number2
StatePublished - 1987

Fingerprint

Steroid Receptors
Steroids
Cytoplasmic and Nuclear Receptors
Oviducts
Avidin
Cell Separation
Charcoal
Progesterone Receptors
Durapatite
Endometrial Neoplasms
Progesterone
Breast Neoplasms
Temperature
Proteins

ASJC Scopus subject areas

  • Endocrinology
  • Endocrinology, Diabetes and Metabolism

Cite this

Spelsberg, T. C., Graham, M. L., Berg, N. J., Umehara, T., Riehl, E., Coulam, C. B., & Ingle, J. N. (1987). A nuclear binding assay to assess the biological activity of steroid receptors in isolated animal and human tissues. Endocrinology, 121(2), 631-644.

A nuclear binding assay to assess the biological activity of steroid receptors in isolated animal and human tissues. / Spelsberg, T. C.; Graham, M. L.; Berg, N. J.; Umehara, T.; Riehl, E.; Coulam, C. B.; Ingle, J. N.

In: Endocrinology, Vol. 121, No. 2, 1987, p. 631-644.

Research output: Contribution to journalArticle

Spelsberg, TC, Graham, ML, Berg, NJ, Umehara, T, Riehl, E, Coulam, CB & Ingle, JN 1987, 'A nuclear binding assay to assess the biological activity of steroid receptors in isolated animal and human tissues', Endocrinology, vol. 121, no. 2, pp. 631-644.
Spelsberg TC, Graham ML, Berg NJ, Umehara T, Riehl E, Coulam CB et al. A nuclear binding assay to assess the biological activity of steroid receptors in isolated animal and human tissues. Endocrinology. 1987;121(2):631-644.
Spelsberg, T. C. ; Graham, M. L. ; Berg, N. J. ; Umehara, T. ; Riehl, E. ; Coulam, C. B. ; Ingle, J. N. / A nuclear binding assay to assess the biological activity of steroid receptors in isolated animal and human tissues. In: Endocrinology. 1987 ; Vol. 121, No. 2. pp. 631-644.
@article{97d11023103e49b2b3d9e006a5ef7d63,
title = "A nuclear binding assay to assess the biological activity of steroid receptors in isolated animal and human tissues",
abstract = "This paper describes a nuclear binding assay (NB assay) which measures not only the presence of a steroid receptor in a tissue, but also the quantity of that receptor which is biologically active or functional, i.e. able to bind to nuclear acceptor sites. The assay involes the isolation viable cells from tissues and their incubation with an excess of radiolabeled steroid to encourage the activation and nuclear binding of all cellular receptors. The nuclei are isolated under conditions that remove unactivated (unbound) steroid-receptor complexes. This NB assay demonstrates, in both animal and human steroid target tissues, a saturable, tissue- and steroid-specific, and temperature- and time-dependent nuclear binding of radiolabeled steroids. These properties support a receptor-dependent nuclear binding of steroids. This assay is reproducible and requires relatively small amounts of tissue. The patterns of nuclear binding of the progesterone receptor, achieved with the assay in the avian oviduct model system, are shown to correlate with the nuclear binding of progesterone in vivo, the ability of the steroid to alter transcription, and the expression of a specific gene product, the protein avidin. The assay has been used to identify the existence of nonfunctional steroid receptors in endometrial and breast carcinomas. Therefore, this NB assay combined with the standard charcoal/hydroxylapatite methods of quantitating total cellular receptors should provide a means of assessing changes in the regulation of the biological activity of steroid receptors. Further, the assay should be useful to assess the ability of steroid analogs to properly activate their respective receptors for subsequent nuclear binding.",
author = "Spelsberg, {T. C.} and Graham, {M. L.} and Berg, {N. J.} and T. Umehara and E. Riehl and Coulam, {C. B.} and Ingle, {J. N.}",
year = "1987",
language = "English (US)",
volume = "121",
pages = "631--644",
journal = "Endocrinology",
issn = "0013-7227",
publisher = "The Endocrine Society",
number = "2",

}

TY - JOUR

T1 - A nuclear binding assay to assess the biological activity of steroid receptors in isolated animal and human tissues

AU - Spelsberg, T. C.

AU - Graham, M. L.

AU - Berg, N. J.

AU - Umehara, T.

AU - Riehl, E.

AU - Coulam, C. B.

AU - Ingle, J. N.

PY - 1987

Y1 - 1987

N2 - This paper describes a nuclear binding assay (NB assay) which measures not only the presence of a steroid receptor in a tissue, but also the quantity of that receptor which is biologically active or functional, i.e. able to bind to nuclear acceptor sites. The assay involes the isolation viable cells from tissues and their incubation with an excess of radiolabeled steroid to encourage the activation and nuclear binding of all cellular receptors. The nuclei are isolated under conditions that remove unactivated (unbound) steroid-receptor complexes. This NB assay demonstrates, in both animal and human steroid target tissues, a saturable, tissue- and steroid-specific, and temperature- and time-dependent nuclear binding of radiolabeled steroids. These properties support a receptor-dependent nuclear binding of steroids. This assay is reproducible and requires relatively small amounts of tissue. The patterns of nuclear binding of the progesterone receptor, achieved with the assay in the avian oviduct model system, are shown to correlate with the nuclear binding of progesterone in vivo, the ability of the steroid to alter transcription, and the expression of a specific gene product, the protein avidin. The assay has been used to identify the existence of nonfunctional steroid receptors in endometrial and breast carcinomas. Therefore, this NB assay combined with the standard charcoal/hydroxylapatite methods of quantitating total cellular receptors should provide a means of assessing changes in the regulation of the biological activity of steroid receptors. Further, the assay should be useful to assess the ability of steroid analogs to properly activate their respective receptors for subsequent nuclear binding.

AB - This paper describes a nuclear binding assay (NB assay) which measures not only the presence of a steroid receptor in a tissue, but also the quantity of that receptor which is biologically active or functional, i.e. able to bind to nuclear acceptor sites. The assay involes the isolation viable cells from tissues and their incubation with an excess of radiolabeled steroid to encourage the activation and nuclear binding of all cellular receptors. The nuclei are isolated under conditions that remove unactivated (unbound) steroid-receptor complexes. This NB assay demonstrates, in both animal and human steroid target tissues, a saturable, tissue- and steroid-specific, and temperature- and time-dependent nuclear binding of radiolabeled steroids. These properties support a receptor-dependent nuclear binding of steroids. This assay is reproducible and requires relatively small amounts of tissue. The patterns of nuclear binding of the progesterone receptor, achieved with the assay in the avian oviduct model system, are shown to correlate with the nuclear binding of progesterone in vivo, the ability of the steroid to alter transcription, and the expression of a specific gene product, the protein avidin. The assay has been used to identify the existence of nonfunctional steroid receptors in endometrial and breast carcinomas. Therefore, this NB assay combined with the standard charcoal/hydroxylapatite methods of quantitating total cellular receptors should provide a means of assessing changes in the regulation of the biological activity of steroid receptors. Further, the assay should be useful to assess the ability of steroid analogs to properly activate their respective receptors for subsequent nuclear binding.

UR - http://www.scopus.com/inward/record.url?scp=0023219563&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0023219563&partnerID=8YFLogxK

M3 - Article

VL - 121

SP - 631

EP - 644

JO - Endocrinology

JF - Endocrinology

SN - 0013-7227

IS - 2

ER -