Properties and interpretation of the woolf and scatchard plots in analysing data from steroid receptor assays

Keightley, D.D.; Fisher, Robert J.; Cressie, Noel

doi:10.1016/0022-4731(83)91115-9

Cited by 48 publications

(13 citation statements)

References 13 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Binding equilibrium was reached after incubation for 3 h at 0°C. Separation of bound and free 3H-labelled steroid was performed by Sephadex LH-20 gel filtration (de Kloet et al 1975 (Haldane, 1957;Keightley & Cressie, 1980;Keightley, Fisher & Cressie, 1983) and Scatchard (Scatchard, 1949) analysis respectively. Figure 1 shows the distribution of type-I and type-II receptors in rat brain, as measured by quantitative image analysis following in-vitro autoradiography.…”

Section: Animalsmentioning

confidence: 99%

Relative occupation of type-I and type-II corticosteroid receptors in rat brain following stress and dexamethasone treatment: functional implications

Reul¹,

Bosch²,

Kloet³

1987

Journal of Endocrinology

367

215

View full text Add to dashboard Cite

The rat brain contains two receptor systems for corticosterone: the type-I corticosterone-preferring receptor and the classical type-II glucocorticoid receptor. The two receptor populations can be distinguished in binding studies with the 'pure' synthetic glucocorticoid 11 beta,17 beta-dihydroxy-6-methyl-17 alpha (1-propynyl)-androsta-1,4,6-trione-3-one (RU 28362). In-vitro autoradiography and quantitative image analysis showed that the type-I receptor was localized almost exclusively in the hippocampus, whereas the type-II receptor extended throughout the brain, with the highest levels in the nucleus paraventricularis, nucleus supraopticus and in the thalamic, amygdaloid, hippocampal and septal regions. Unoccupied type-I and type-II receptor sites, as measured in vitro by cytosol binding of 3H-labelled steroids, displayed a large difference in the rate of appearance after adrenalectomy. The availability of type-I receptors exhibited a marked increase, reaching maximal levels within 4-7 h, and then remained constant until 2 weeks after adrenalectomy. The availability of type-II receptors did not change considerably during the first 24 h after adrenalectomy, but displayed a large increase in capacity during the subsequent 2 weeks. After adrenocortical activation as a consequence of exposure to a novel environment, plasma concentrations of corticosterone increased to reach a peak of 811 nmol/l after 30 min and attained the basal concentration (43 nmol/l) after 240 min. During this time, occupation of type-I receptors increased from 77.8% at 0 min to 97% at 30-60 min and then declined to 84.8% after 240 min. Occupation of the type-II receptors was 28.1% at 0 min, 74.5% after 30 min and 32.8% after 240 min.(ABSTRACT TRUNCATED AT 250 WORDS)

show abstract

Section: Animalsmentioning

confidence: 99%

Relative occupation of type-I and type-II corticosteroid receptors in rat brain following stress and dexamethasone treatment: functional implications

Reul¹,

Bosch²,

Kloet³

1987

Journal of Endocrinology

367

215

View full text Add to dashboard Cite

show abstract

“…15 The apo B/E receptormediated binding was determined by subtracting nonspecific from total binding, and the binding parameters K d and B^ were determined from Woolf plots. 23 …”

Section: In Vitro Ldl Bindingmentioning

confidence: 99%

Dietary fat saturation modifies the metabolism of LDL subfractions in guinea pigs.

Fernández

Abdel-Fattah

McNamara

1993

Arterioscler Thromb

View full text Add to dashboard Cite

The effects of dietary fat saturation on the metabolism of low-density Iipoprotein (LDL) subtractions were measured in adult male guinea pigs fed semipurified diets containing 15% (wt/wt) corn oil (CO; 58% llnoleic acid), lard (24% palmitic/14% stearic acid), or palm kernel oil (PK; 52% lauric/18% myristic acid). Animals fed the CO diet had lower plasma total cholesterol levels than guinea pigs fed the PK or lard diets (P<.01). Plasma LDL-1 (<.01). Within the same diet group, LDL-2 exhibited a slower turnover rate than LDL-1 in animals fed the PK diet, while no differences in LDL subtraction FCR were found in the CO and lard groups. Animals fed the PK and lard diets did not exhibit significant modifications in the density distribution of LDL subtractions over a period of 33 hours. In contrast, animals fed the CO diet exhibited a shift of more buoyant to denser LDL particles, suggesting that differences in LDL intravascular processing are mediated by dietary fat saturation. In vitro LDL binding to hepatic membranes confirmed the in vivo data with an increased expression of apolipoprotein B/E receptors ( B m ) in animals fed the CO diet (P<.01). Hepatic apolipoprotein B/E receptors exhibited less affinity for LDL-2 in the PK group, a result consistent with the less rapid turnover of LDL-2 in PK-fed animals. The results suggest that dietary fatty acids varying in saturation and composition have distinctive atherogenic potentials. The lowest plasma LDL cholesterol concentrations mediated by CO intake could in part be explained by induced changes in the composition and processing of LDL subtractions, resulting in faster LDL turnover rates in addition to increased expression of hepatic apolipoprotein B/E receptors. Similarly, the hypercholesterolemic effects of the lard and PK diets appear to be related to differences in intravascular processing of LDL subtractions, with no interconversion between LDL-1 and LDL-2 and that is accompanied by decreased expression of hepatic apo B/E receptors. In addition, PK-fed guinea pigs exhibited a decreased apolipoprotein B/E receptor affinity for LDL-2. {ArterioscUr Thromb. 1993;13:1418-1428.) KEY WORDS • LDL subtractions • dietary fat saturation • catabolic rates • apo B/E receptor • guinea pigs plasma LDL distribution • fractionalN umerous studies have investigated the effects of dietary fatty acids on plasma low-density lipoprotein (LDL) transport to determine the mechanisms by which specific fatty acids alter plasma LDL levels. 1 Studies of LDL kinetics have...

show abstract

“…Data were expressed as femtomoles/per milligram protein. The apparent maximal binding ca pacity (Bma,i) for CR and GR was calculated by Woolf analysis [10].…”

Section: Binding Assaysmentioning

confidence: 99%

Differential Response of Type I and Type II Corticosteroid Receptors to Changes in Plasma Steroid Level and Circadian Rhythmicity

Reul

Bosch

Kloet³

1987

Neuroendocrinology

250

147

View full text Add to dashboard Cite

Corticosterone (CORT) binds to two receptor systems in rat brain: the type I CORT-preferring receptor (CR) and the type II glucocorticoid receptor (GR). Discrimination between the two receptor types can be achieved with the ‘pure’ synthetic glucocorticoid RU 28362. In this study, we show that the binding capacity of GR in the rat hippocampus exhibits a strikingly different response from CR to adrenalectomy (ADX), chronic steroid replacement, hypophysectomy (HYPOX) and during circadian variation. Under those experimental conditions neither receptor site showed changes in binding affinity. After ADX, CR number remained relatively constant for a period of 13 days, while GR capacity increased by 133%, a level which was reached 5 days post-surgery. CR capacity showed circadian variation, since CR number was 65% higher in the evening than in the morning. GR capacities at those two time points were not significantly different. Replacement with subcutaneous CORT implants (100-mg pellets) for 7 days following ADX rats did not affect CR number, but caused a 38% decrease in GR number compared to control animals (cholesterol-treated, 7-day-ADX rats). On the other hand, dexamethasone (DEX) implants (5-, 15-, 25-mg pellets) elicited a dose-dependent increase in CR capacity (up to 99%) and a dose-dependent decrease in GR capacity (40–44%). Finally, 2 weeks after HYPOX, CR and GR numbers were increased by 60 and 38%, respectively. We conclude that the type II GR capacity responds in an autoregulatory manner to changes in circulating plasma glucocorticoid levels, while type I CR does not.

show abstract

Properties and interpretation of the woolf and scatchard plots in analysing data from steroid receptor assays

Cited by 48 publications

References 13 publications

Relative occupation of type-I and type-II corticosteroid receptors in rat brain following stress and dexamethasone treatment: functional implications

Relative occupation of type-I and type-II corticosteroid receptors in rat brain following stress and dexamethasone treatment: functional implications

Dietary fat saturation modifies the metabolism of LDL subfractions in guinea pigs.

Differential Response of Type I and Type II Corticosteroid Receptors to Changes in Plasma Steroid Level and Circadian Rhythmicity

Contact Info

Product

Resources

About