Glucocorticoid receptor in circulating mononuclear leukocytes

Tohru Murakami, David Brandon, David Rodbard, D. Lynn Loriaux, Mortimer B. Lipsett

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31 Scopus citations


The hypothesis that 17β-estradiol suppresses dopamine secretion into hypophysial portal blood was tested. Portal plasma concentrations of dopamine were significantly lower in proestrous rats (1.0 ± 0.1 ng/ml; mean ± SE) than in estrous rats (1.9 ± 0.38 ng/ml). To deplete the animal of endogenous steroid hormones, proestrous rats were adrenalectomized (Adx) and ovariectomized (Ovx). Twenty-four hours later, hypophysial portal blood was collected for 60 min, and the plasma from this blood was analyzed for dopamine. Arterial plasma from these rats was assayed for 17β-estradiol and progesterone. The concentrations of dopamine in the portal plasma of sham-operated rats and bilaterally Adx-Ovx rats were similar to those in estrous animals. The concentration of dopamine in portal plasma of Adx-Ovx rats injected 24 h earlier with 50 μg 17β-estradiol was 1.0 ± 0.31 ng/ml, which was comparabExposure of adipocytes from young rats (2–3 months old) to dexamethasone in vitro results in 40–50% inhibition of glucose transport and metabolism. ComparabDose-response curves were obtained for the production of androgen-binding protein (ABP) by Sertoli cells prepared from testes of 20-day-old rats and treated in culture with either FSH or testosterone (T). FSH stimulated ABP production by up to 3.5 times control levels. For NIH-FSH-Sll, the ED50 was 3 ng/ml, and for highly purified ovine FSH, the ED50 was 0.066 ng/ml. Addition of T produced a stimulation of up to 3 times control levels; half-maximal response was obtained at a dose of 4 nM. The presence of small numbers of contaminating Leydig cells in some preparations resulted in production of endogenous T, especially when high doses of NIH-FSH, which contains some LH, were employed. A modified preparatDissociation of [125I]iodoinsulin from adipocyte insulin receptors was studied in the presence or absence of the insulin derivatives, desoctapeptide insulin and desalanine desasparagine insulin. When cells were allowed to associate with a tracer concentration (10-10 M) of [125I]iodoinsulin and dissociation was studied in either insulin-free buffer or buffer containing 100 ng/ml unlabeled insulin, dissociation was accelerated in the presence of unlabeled insulin. This is consistent with negatively cooperative site-site interactions. On the other hand, when dissociation studies were performed in the presence of high concentrations of desoctapeptide insulin or desalanine desasparagine insulin, dissociation rates were slower than those observed in insulin-free buffer. In marked contrast, when cells were allowed to achieve a high fractional receptor occupancy by associating with high concentrations of either desoctapeptide insulin or desalanine desasparagTo study the relationship between formation and release of Golgi-derived secretory granules and progesterone secretion, slices of ovine luteal tissue were incubated in the presence of LH and/or calcium ionophore A23187. Increases in progesterone secretion in response to LH and/or ionophore were accompanied by a concomitant release of secretory granules. In contrast, in the presence of colchicine, LH-stimulated progesterone secretion was significantly reduced (P < 0.01), granule formation appeared to be blocked, and there was little evidence of exocytosis. In addition, unusual pleomorphic membranebounded saccules containing an electron-dense material were abundant throughout the centrospheric region of cells treated with colchicine. Because of the close paralEffects of ionophore A23187 on skeletal collagen formation were investigated in vitro. Collagen synthesis was quantitated in fetal rat calvaria by measuring [3H]proline incorporation into collagenase-digestible (CDP) and noncollagen protein (NCP) using purified bacterial collagenase; [3H]proline was added for the last 2 h of culture. Results are as follows. 1) A23187 (0.03–1.0 jug:/ml) inhibited incorporation of label into CDP and NCP after 24 h of culture, with a greater effect on CDP. The response was not associated with altered amino acid uptake, precursor pool size, or degradation of newly labeled protein. 2) Submaximal concentrations of A23187 and parathyroid hormone or dibutyryl cAMP decreased CDP formation to a greater extent than treatment with the agents alone. 3) Imidazole, while ineffective by itself, enhanced the effect of A23187. 4) Alteration of medium calcium did not affect the response to ionophore. 5) The inhibitory effect of A23187 was partially reversed by 24 h and completely reversed by 48 h of control treatment subsequent to an initial 24-h incubation with ionophore. 6) Indomethacin had no effect on CDP or NCP formation, either in the presence or absence of A23187. 7) A23187 did not alter the uptake of [3H]thymidine or [3H]uridine into acid-extractable pools but decreased incorporation of label into DNA and RNA, respectively. 8) Histological examination showed no difference between control and A23187 treatment after 24 h. We conclude that A23187 decreases bone collagen and noncollagen protein synthesis, possibly through a calcium-mediated effect. The mechanism of the inhibitory effect on DNA and RNA labeling is unknown, although it may be related to calcium. Our results further suggest that calcium may be involved in the actions of paratBinding of [3H]dexamethasone ([3H]Dex) and [3H]cortisol to normal female goat mononuclear leukocytes (MNL) was studied at 4 and 20 C. Dissociation studies of [3H]Dex and [3H]cortisol from MNL after 3 h of incubation at 20 C indicated the presence of at least two components; a small pool with a dissociation rate constant (kOfr) of 15 ± 8 × 10-3 min-1 for [3H]Dex and 35 ± 11 × 10-3 min-1 for [3H]cortisol, and a large capacity pool with a kOff of 1.0 ± 0.1 × 10-3 min-1 for [3H]Dex and 11 ± 2 × 10-3 min-1 for [3H]cortisol. The calculated koff for the slow component was indistinguishable from the kOfr for nuclei isolated from cells that had been preincubated with [3H]Dex or [3H]cortisol. Thus, for the intact cells the slow component probably represents the dissociation of an activated steroid-receptor complex transformed by a temperature-dependent process at 20 C. The rapid phase may represent the dissociation of an inactive steroid-receptor complex in the cytosol at 20 C. The Kd obtained from equilibrium Scatchard analysis (Dex, 0.27 ± 0.02 nM; cortisol, 4.5 ± 0.1 nM) was in satisfactory agreement with the ratio of kOfr to association rate from kinetic studies. The dissociation of [3H]Dex and [3H]cortisol from MNL at 4 C can be described by a single exponential decay. The ratio of koff tp association rate at 4 C corresponds to the KdS obtained using equilibrium Scatchard analysis (at 15 h) for either intact cells (Dex, 2.0 ± 0.3 nM; cortisol, 8.0 ± 0.9 nM) or cytosols (Dex, 1.4 ± 0.4 nM; cortisol, 8.5 ± 1.9 nM). The higher affinity of Dex to the intact cell at 20 C may explain the previously noted discrepancy between the increased biological potency of Dex compared to that of cortisol and the modest increase in binding affinity of Dex to cytosol receptors.

Original languageEnglish (US)
Pages (from-to)500-505
Number of pages6
Issue number2
StatePublished - Feb 1979


ASJC Scopus subject areas

  • Endocrinology

Cite this

Murakami, T., Brandon, D., Rodbard, D., Loriaux, D. L., & Lipsett, M. B. (1979). Glucocorticoid receptor in circulating mononuclear leukocytes. Endocrinology, 104(2), 500-505.