Changes in [125I]labeled human chorionic gonadotropin (hCG) binding to porcine granulosa cells during follicle development and cell culture

Richard Stouffer, L. Tyrey, D. W. Schomberg

Research output: Contribution to journalArticle

25 Citations (Scopus)

Abstract

The specific binding of [125I]iodo human chorionic gonadotropin to porcine granulosa cells isolated at 2 stages of follicle maturation was quantitated immediately after harvest and following 6 to 7 days of culture. Both porcine LH (pLH, LER 786-3) and hCG inhibited [125I]iodo hCG binding, while pFSH (LER 1132) did not compete for hCG binding sites. The incubation of labeled hCG with granulosa cells for 24 to 48 hr at 37°C did not alter its subsequent ability to bind to fresh cells. The binding of [125I]iodo hCG was a rapid process which was not readily reversible. Scatchard analysis of the equilibrium binding data resulted in linear plots showing the hCG binding capacity of highly differentiated (HD) cells, harvested from large preovulatory follicles, to be significantly greater than that of moderately differentiated (MD) cells isolated from smaller luteal phase follicles, both before (794 vs. 93 pmoles/g; P <0.001) and after (157 vs. 5 pmoles/g; P <0.001) culture. A decline in binding capacity occurred in both cell groups during culture and was associated with a significant decrease in progestin production. In contrast, the hCG binding affinity of the granulosa cell was equivalent at both stages of follicle development and remained unchanged after 6 days of culture (mean apparent Kd = 2.9 x 10-10M; n = 27). These data indicate that porcine granulosa cells contain a homogeneous class of LH receptors whose number, but not affinity, increases during follicle maturation in vivo. The loss of receptors in vitro suggests the absence of some factor(s), as yet unidentified, critical to the maintenance and development of the receptor population.

Original languageEnglish (US)
Pages (from-to)516-525
Number of pages10
JournalEndocrinology
Volume99
Issue number2
StatePublished - 1976
Externally publishedYes

Fingerprint

Granulosa Cells
Chorionic Gonadotropin
Swine
Cell Culture Techniques
LH Receptors
Luteal Phase
Progestins
Binding Sites
Maintenance

ASJC Scopus subject areas

  • Endocrinology
  • Endocrinology, Diabetes and Metabolism

Cite this

Changes in [125I]labeled human chorionic gonadotropin (hCG) binding to porcine granulosa cells during follicle development and cell culture. / Stouffer, Richard; Tyrey, L.; Schomberg, D. W.

In: Endocrinology, Vol. 99, No. 2, 1976, p. 516-525.

Research output: Contribution to journalArticle

@article{33316e18f0114db7a05dc1d7dd6999ed,
title = "Changes in [125I]labeled human chorionic gonadotropin (hCG) binding to porcine granulosa cells during follicle development and cell culture",
abstract = "The specific binding of [125I]iodo human chorionic gonadotropin to porcine granulosa cells isolated at 2 stages of follicle maturation was quantitated immediately after harvest and following 6 to 7 days of culture. Both porcine LH (pLH, LER 786-3) and hCG inhibited [125I]iodo hCG binding, while pFSH (LER 1132) did not compete for hCG binding sites. The incubation of labeled hCG with granulosa cells for 24 to 48 hr at 37°C did not alter its subsequent ability to bind to fresh cells. The binding of [125I]iodo hCG was a rapid process which was not readily reversible. Scatchard analysis of the equilibrium binding data resulted in linear plots showing the hCG binding capacity of highly differentiated (HD) cells, harvested from large preovulatory follicles, to be significantly greater than that of moderately differentiated (MD) cells isolated from smaller luteal phase follicles, both before (794 vs. 93 pmoles/g; P <0.001) and after (157 vs. 5 pmoles/g; P <0.001) culture. A decline in binding capacity occurred in both cell groups during culture and was associated with a significant decrease in progestin production. In contrast, the hCG binding affinity of the granulosa cell was equivalent at both stages of follicle development and remained unchanged after 6 days of culture (mean apparent Kd = 2.9 x 10-10M; n = 27). These data indicate that porcine granulosa cells contain a homogeneous class of LH receptors whose number, but not affinity, increases during follicle maturation in vivo. The loss of receptors in vitro suggests the absence of some factor(s), as yet unidentified, critical to the maintenance and development of the receptor population.",
author = "Richard Stouffer and L. Tyrey and Schomberg, {D. W.}",
year = "1976",
language = "English (US)",
volume = "99",
pages = "516--525",
journal = "Endocrinology",
issn = "0013-7227",
publisher = "The Endocrine Society",
number = "2",

}

TY - JOUR

T1 - Changes in [125I]labeled human chorionic gonadotropin (hCG) binding to porcine granulosa cells during follicle development and cell culture

AU - Stouffer, Richard

AU - Tyrey, L.

AU - Schomberg, D. W.

PY - 1976

Y1 - 1976

N2 - The specific binding of [125I]iodo human chorionic gonadotropin to porcine granulosa cells isolated at 2 stages of follicle maturation was quantitated immediately after harvest and following 6 to 7 days of culture. Both porcine LH (pLH, LER 786-3) and hCG inhibited [125I]iodo hCG binding, while pFSH (LER 1132) did not compete for hCG binding sites. The incubation of labeled hCG with granulosa cells for 24 to 48 hr at 37°C did not alter its subsequent ability to bind to fresh cells. The binding of [125I]iodo hCG was a rapid process which was not readily reversible. Scatchard analysis of the equilibrium binding data resulted in linear plots showing the hCG binding capacity of highly differentiated (HD) cells, harvested from large preovulatory follicles, to be significantly greater than that of moderately differentiated (MD) cells isolated from smaller luteal phase follicles, both before (794 vs. 93 pmoles/g; P <0.001) and after (157 vs. 5 pmoles/g; P <0.001) culture. A decline in binding capacity occurred in both cell groups during culture and was associated with a significant decrease in progestin production. In contrast, the hCG binding affinity of the granulosa cell was equivalent at both stages of follicle development and remained unchanged after 6 days of culture (mean apparent Kd = 2.9 x 10-10M; n = 27). These data indicate that porcine granulosa cells contain a homogeneous class of LH receptors whose number, but not affinity, increases during follicle maturation in vivo. The loss of receptors in vitro suggests the absence of some factor(s), as yet unidentified, critical to the maintenance and development of the receptor population.

AB - The specific binding of [125I]iodo human chorionic gonadotropin to porcine granulosa cells isolated at 2 stages of follicle maturation was quantitated immediately after harvest and following 6 to 7 days of culture. Both porcine LH (pLH, LER 786-3) and hCG inhibited [125I]iodo hCG binding, while pFSH (LER 1132) did not compete for hCG binding sites. The incubation of labeled hCG with granulosa cells for 24 to 48 hr at 37°C did not alter its subsequent ability to bind to fresh cells. The binding of [125I]iodo hCG was a rapid process which was not readily reversible. Scatchard analysis of the equilibrium binding data resulted in linear plots showing the hCG binding capacity of highly differentiated (HD) cells, harvested from large preovulatory follicles, to be significantly greater than that of moderately differentiated (MD) cells isolated from smaller luteal phase follicles, both before (794 vs. 93 pmoles/g; P <0.001) and after (157 vs. 5 pmoles/g; P <0.001) culture. A decline in binding capacity occurred in both cell groups during culture and was associated with a significant decrease in progestin production. In contrast, the hCG binding affinity of the granulosa cell was equivalent at both stages of follicle development and remained unchanged after 6 days of culture (mean apparent Kd = 2.9 x 10-10M; n = 27). These data indicate that porcine granulosa cells contain a homogeneous class of LH receptors whose number, but not affinity, increases during follicle maturation in vivo. The loss of receptors in vitro suggests the absence of some factor(s), as yet unidentified, critical to the maintenance and development of the receptor population.

UR - http://www.scopus.com/inward/record.url?scp=0017098399&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0017098399&partnerID=8YFLogxK

M3 - Article

VL - 99

SP - 516

EP - 525

JO - Endocrinology

JF - Endocrinology

SN - 0013-7227

IS - 2

ER -