Changes in Cardiac and Hepatic Glucocorticoid Receptors after Adrenalectomy

1976 ◽  
Vol 51 (5) ◽  
pp. 487-493
Author(s):  
M. C. Gregory ◽  
D. Duval ◽  
P. Meyer
Keyword(s):  
Intravenous Injection ◽  
Binding Sites ◽  
Rat Heart ◽  
Glucocorticoid Receptors ◽  
Liver Cytosol ◽  
Receptor Complexes ◽  
Number Of Binding Sites ◽  
Endogenous Steroids

1. The total number of specific dexamethasone-binding sites in rat heart and liver cytosol was measured at intervals after adrenalectomy. 2. Between 12 and 48 h after adrenalectomy there was a significant increase in the number of binding sites in both heart and liver cytosol. Affinity was unchanged. 3. Of the [3H]corticosterone bound to liver cytosol proteins after an intravenous injection 98% disappeared within 2 h in vivo. Dissociation of endogenous corticosterone-receptor complexes in liver cytosol will thus be substantially complete some hours before the number of receptors increases. 4. It was concluded that there is a true increase in the number of glucocorticoid receptors occurring principally between 12 and 48 h after suppression of endogenous steroids.

scholarly journals Two Populations of Glucocorticoid Receptor-Binding Sites in the Male Rat Hippocampal Genome

Endocrinology ◽  
2013 ◽  
Vol 154 (5) ◽  
pp. 1832-1844 ◽  
Author(s):  
J. Annelies E. Polman ◽  
E. Ronald de Kloet ◽  
Nicole A. Datson
Keyword(s):  
Receptor Binding ◽  
Binding Sites ◽  
Glucocorticoid Receptors ◽  
Full Range ◽  
Male Rat ◽  
Motif Analysis ◽  
Wide Range ◽  
Neuronal Processes ◽  
Almost All

Abstract In the present study, genomic binding sites of glucocorticoid receptors (GR) were identified in vivo in the rat hippocampus applying chromatin immunoprecipitation followed by next-generation sequencing. We identified 2470 significant GR-binding sites (GBS) and were able to confirm GR binding to a random selection of these GBS covering a wide range of P values. Analysis of the genomic distribution of the significant GBS revealed a high prevalence of intragenic GBS. Gene ontology clusters involved in neuronal plasticity and other essential neuronal processes were overrepresented among the genes harboring a GBS or located in the vicinity of a GBS. Male adrenalectomized rats were challenged with increasing doses of the GR agonist corticosterone (CORT) ranging from 3 to 3000 μg/kg, resulting in clear differences in the GR-binding profile to individual GBS. Two groups of GBS could be distinguished: a low-CORT group that displayed GR binding across the full range of CORT concentrations, and a second high-CORT group that displayed significant GR binding only after administering the highest concentration of CORT. All validated GBS, in both the low-CORT and high-CORT groups, displayed mineralocorticoid receptor binding, which remained relatively constant from 30 μg/kg CORT upward. Motif analysis revealed that almost all GBS contained a glucocorticoid response element resembling the consensus motif in literature. In addition, motifs corresponding with new potential GR-interacting proteins were identified, such as zinc finger and BTB domain containing 3 (Zbtb3) and CUP (CG11181 gene product from transcript CG11181-RB), which may be involved in GR-dependent transactivation and transrepression, respectively. In conclusion, our results highlight the existence of 2 populations of GBS in the rat hippocampal genome.

Endothelin ETA and ETB receptors in postnatal intestine

2001 ◽  
Vol 280 (4) ◽  
pp. G555-G562 ◽  
Author(s):  
Craig A. Nankervis ◽  
David J. Dunaway ◽  
Charles E. Miller
Keyword(s):  
Binding Sites ◽  
Specific Binding ◽  
Ischemia Reperfusion ◽  
Age Groups ◽  
Scatchard Analysis ◽  
Binding Assays ◽  
Site Model ◽  
Number Of Binding Sites ◽  
Competitive Binding Assays

We aimed to characterize endothelin (ET) receptors in the swine intestinal vasculature and to determine ischemia-reperfusion (I/R) effects on these receptors. Saturation and competitive binding assays were performed on mesenteric artery protein membranes from 1- and 40-day-old animals, both control and those subjected to 1 h of partial ischemia followed by 6 h of reperfusion in vivo. Scatchard analysis of saturation binding with 125I-labeled ET-1 in membranes from endothelium-denuded (E−) vessels revealed that the maximum number of binding sites was greater in younger animals. Competitive125I-ET-1 binding was significant for a one-site model with ET-1, ET-3, and sarafotoxin S6c (S6c) in membranes from endothelium-intact (E+) and E− vessels in both age groups. The maximum number of ET-1 binding sites was significantly greater in younger animals. In the presence of the ETAreceptor antagonist BQ-123, competitive 125I-ET-1 binding was significant for a one-site model with ET-1 and S6c in membranes from E+ vessels in both age groups. The maximum number of ET-1 binding sites was significantly greater in younger animals. After I/R, the maximum number of ET-1 binding sites was unchanged. In the presence of BQ-123, specific binding by ET-1 and S6c was eliminated in both age groups after I/R. These results suggest that both ET receptor populations are expressed to a greater degree in younger animals and I/R significantly affects the ETB receptor.

scholarly journals The binding of spermine to the ribosomes and ribosomal ribonucleic acid from Bacillus stearothermophilus

1969 ◽  
Vol 113 (1) ◽  
pp. 117-121 ◽  
Author(s):  
L. Stevens
Keyword(s):  
Ionic Strength ◽  
Ribosomal Rna ◽  
Ribonucleic Acid ◽  
Binding Sites ◽  
Bacillus Stearothermophilus ◽  
Gel Filtration ◽  
Ribosomal Ribonucleic Acid ◽  
Number Of Binding Sites ◽  
Filtration Technique

1. The total intracellular concentrations of Na+, K+, Mg2+, spermine, spermidine and RNA were measured in Bacillus stearothermophilus. 2. The binding of spermine to ribosomes and to ribosomal RNA from B. stearothermophilus was studied under various conditions by using a gel-filtration technique. 3. The affinity of spermine for ribosomes and for ribosomal RNA decreased with increasing ionic strength of the medium in which they were suspended. 4. The extent of spermine binding did not change appreciably in the temperature range 4–60°. 5. Optimum binding occurred at about pH7·0. 6. The number of binding sites for spermine on either ribosomes or ribosomal RNA was 0·10–0·13/RNA phosphate group. 7. A high proportion of the intracellular spermine is likely to be bound to the ribosomes in vivo; spermine competes with Mg2+ on equal terms for sites on the ribosomes.

The interaction of sialidase-treated hCG with ovarian cellular membranes

1981 ◽  
Vol 97 (2) ◽  
pp. 270-280 ◽  
Author(s):  
E. C. Brand ◽  
J. Odink ◽  
G. Klok ◽  
E. V. van Hall
Keyword(s):  
Binding Sites ◽  
Association Constant ◽  
Apparent Difference ◽  
Receptor Complexes ◽  
Number Of Binding Sites ◽  
Biological Potency ◽  
The Difference ◽  
The Stability ◽  
Dissociation Curves ◽  
Hcg Receptor

Abstract. The potency of human chorionic gonadotrophin (hCG) in competition for binding to a gonadal membrane fraction is remarkably enhanced by sialidase treatment. The present study was undertaken to investigate the specificity and characteristics of the binding of sialidase-treated hCG (asialo-hCG) in a particulate hCG-binding system from luteinized rat ovaries. The competitive potency of asialo-hCG relative to hCG was 2.5, irrespective of whether [125I]hCG or [125I]asialo-hCG was used for tracer. This was due to a 2.1 times higher equilibrium association constant for asialo-hCG, whereas the estimated number of binding sites did not differ. There was no apparent difference in the stability of hCG and asialo-hCG, or in the stability of the respective hormone-receptor complexes. The effect of variation of the incubation conditions on the binding of both tracers was similar. In accordance with the difference in the equilibrium association constant, the association velocity of asialo-hCG was more than double that of hCG. With all of the tracers used the dissociation curves were biphasic, the size of the initial fast-dissociating fraction being inversely related to the pre-incubation time. Under identical conditions, the fast-dissociating fraction was smaller for the [125I]asialo-hCG complex than for the [125I]hCG complex. The dissociation velocities of these fractions appeared to be similar. The results indicate that asialo-hCG binds to the hCG receptor in a way similar to the binding of the unmodified hormone, but with a higher affinity. The smaller size of the fast-dissociation form of the asialo-hCG-receptor complex may be related to the lower biological potency of the hormone derivative.

Interaction of spironolactone with rat skin androgen receptor

1979 ◽  
Vol 57 (7) ◽  
pp. 1042-1046 ◽  
Author(s):  
Andrée Boisselle ◽  
France T. Dionne ◽  
Roland R. Tremblay
Keyword(s):  
Androgen Receptor ◽  
Binding Sites ◽  
Competitive Inhibition ◽  
Male Rats ◽  
Affinity Constant ◽  
Dorsal Skin ◽  
Rat Skin ◽  
Number Of Binding Sites ◽  
Rat Prostate

Some characteristics of the dorsal skin cytoplasmic androgen receptor (AR) have been studied in male rats. The affinity constant, the binding specificity, and the sedimentation profile of the receptor have been found to be similar to the rat prostate AR. The measurement of the number of binding sites in various hormonal conditions (deprivation) led to the conclusion that this receptor was largely occupied by endogeneous hormones from gonadal and (or) adrenal sources. Administration of spironolactone or canrenone to 7-day-castrated rats was accompanied by a rapid and drastic decrease of available binding sites. This diminution was ascribed to the competitive inhibition of canrenone, the active in vivo metabolite of spironolactone. It is postulated that the antiandrogenic action of spironolactone, at the skin level, is mediated by canrenone which inhibits the formation of specific testosterone and (or) 5α-dihydrotestosterone receptor complex in cytoplasm and consequently in nuclei.

CHANGES IN OESTRADIOL-17β BINDING IN THE HYPOTHALAMI AND PITUITARY GLANDS OF PERSISTENTLY INFERTILE EWES PREVIOUSLY EXPOSED TO OESTROGENIC SUBTERRANEAN CLOVER: EVIDENCE OF ALTERATIONS TO OESTRADIOL RECEPTORS

1978 ◽  
Vol 78 (2) ◽  
pp. 171-177 ◽  
Author(s):  
B. Y. TANG ◽  
N. R. ADAMS
Keyword(s):  
Binding Sites ◽  
Association Constant ◽  
Feedback Regulation ◽  
Subterranean Clover ◽  
Pituitary Glands ◽  
Number Of Binding Sites ◽  
And Control ◽  
Apparent Association

SUMMARY The binding of [3H]oestradiol-17β to the hypothalamus and pituitary gland of cloveraffected permanently infertile and control ovariectomized ewes was compared in vivo and in vitro. When [3H]oestradiol-17β was infused into the carotid artery (10 ng/min), the total homogenate and the nuclear and protamine-precipitable cytosol fractions of hypothalami and pituitary glands from clover-affected ewes bound significantly more [3H]oestradiol than those of the controls. Cytoplasmic oestradiol-17β receptors from the pituitary glands of clover-affected ewes showed a significantly lower apparent association constant and a higher number of binding sites/mg protein in vitro. It is suggested that the hypothalami and pituitary glands of ewes made permanently infertile by oestrogenic clover are less sensitive to feedback regulation of oestradiol-17β at physiological levels.

Glucocorticoid receptors in sheep brain tissues during development

1985 ◽  
Vol 249 (4) ◽  
pp. E345-E349 ◽  
Author(s):  
J. C. Rose ◽  
T. E. Kute ◽  
L. Winkler
Keyword(s):  
Binding Sites ◽  
Glucocorticoid Receptors ◽  
Neonatal Period ◽  
Sucrose Density Gradient ◽  
Ligand Binding Site ◽  
Acth Secretion ◽  
Age Related ◽  
Number Of Binding Sites ◽  
Sheep Brain ◽  
Brain Tissues

In this study we describe changes in the number of glucocorticoid binding sites that occur in cytosols of pituitary, hypothalamic, and hippocampal tissue obtained from fetal, newborn, and adult sheep. We observed specific, saturable binding of [3H]triamcinolone acetonide by cytosols of all three tissues. There were both age-related and tissue-related differences in the number of binding sites observed. By midgestation (70 days) significant quantities of binding sites were present in pituitary and brain tissues, and the number remained relatively constant throughout gestation and in the neonatal period. The number of binding sites declined markedly in cytosols from adult tissues. At all ages there were more binding sites present in pituitary cytosols than in cytosols of hypothalamus and hippocampus. The sucrose density gradient profile of the radioactive ligand-binding site complex is characteristic of a glucocorticoid receptor. Thus, by midgestation at least a portion of the biochemical machinery mediating glucocorticoid effects is present in the fetal pituitary and in specific brain tissues considered important in the modulation of ACTH secretion by corticosteroids.

scholarly journals Modelling cytoskeletal transport by clusters of non-processive molecular motors with limited binding sites

2020 ◽  
Vol 7 (8) ◽  
pp. 200527
Author(s):  
Naruemon Rueangkham ◽  
Ian D. Estabrook ◽  
Rhoda J. Hawkins
Keyword(s):  
Molecular Motors ◽  
Binding Sites ◽  
Intracellular Transport ◽  
Exclusion Process ◽  
Stochastic Simulations ◽  
Published Data ◽  
Analytical Work ◽  
Number Of Binding Sites

Molecular motors are responsible for intracellular transport of a variety of biological cargo. We consider the collective behaviour of a finite number of motors attached on a cargo. We extend previous analytical work on processive motors to the case of non-processive motors, which stochastically bind on and off cytoskeletal filaments with a limited number of binding sites available. Physically, motors attached to a cargo cannot bind anywhere along the filaments, so the number of accessible binding sites on the filament should be limited. Thus, we analytically study the distribution and the velocity of a cluster of non-processive motors with limited number of binding sites. To validate our analytical results and to go beyond the level of detail possible analytically, we perform Monte Carlo latticed based stochastic simulations. In particular, in our simulations, we include sequence preservation of motors performing stepping and binding obeying a simple exclusion process. We find that limiting the number of binding sites reduces the probability of non-processive motors binding but has a relatively small effect on force–velocity relations. Our analytical and stochastic simulation results compare well to published data from in vitro and in vivo experiments.

METABOLISM OF TESTOSTERONE AND 5α-REDUCED ANDROGENS BY THE RABBIT PROSTATE AND EPIDIDYMIS: STUDIES IN VITRO AND IN VIVO

1979 ◽  
Vol 82 (2) ◽  
pp. 207-214 ◽  
Author(s):  
W. D. BOOTH ◽  
R. JONES
Keyword(s):  
Blood Plasma ◽  
Intravenous Injection ◽  
Incubation Medium ◽  
Peripheral Circulation ◽  
Adult Rabbit ◽  
Tissue Preparation ◽  
Endogenous Steroids

SUMMARY The metabolism of radioactively labelled testosterone, 5α-dihydrotestosterone, 5α-androstane-3α,17β-diol and 5α-androstane-3β,17β-diol by the rabbit epididymis and prostate has been investigated in vitro and in vivo. In vitro, the rate of conversion of testosterone to 5α-reduced androgens was low in both glands. Varying the nature of the tissue preparation, age of animal or incubation medium did not improve the situation substantially. However, the rabbit prostate and epididymis metabolized 5α-reduced androgens readily. The prostate was particularly efficient at interconverting 5α-androstane-3α,17β-diol and 5α-dihydrotestosterone, whereas the capacity of the epididymis to carry out this step was much lower. Small amounts of 5α-androstane-3,17-dione and androsterone were also identified. Both glands interconverted 5α-dihydrotestosterone and 5α-androstane-3β,17β-diol to a comparable degree. Following the intravenous injection of 3H-labelled testosterone, significant levels of 3H-labelled 5α-dihydrotestosterone were found in the prostate and epididymis within 30 min. Furthermore, 5α-androstane-3α,17β-diol was detected in both glands. In blood plasma, the ratio of radioactively labelled testosterone: 5α-dihydrotestosterone was 2: 1, i.e. similar to that for endogenous steroids. The intravenous injection of 3H-labelled 5α-androstane-3α-17β-diol gave rise to much higher amounts of 5α-dihydrotestosterone in the prostate than in the epididymis, whereas the reverse was found for the levels of unmetabolized diol. The results indicate that the prostate and epididymis of the adult rabbit differ in their capacity to metabolize 5α-reduced androgens and that both glands depend to a large extent on the relatively high levels of 5α-dihydrotestosterone and 5α-androstanediols present in the peripheral circulation, rather than the metabolism of testosterone in situ.

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