Journal of receptor research最新文献_第10页

In situ assay of hormone-stimulated adenylyl cyclase in 96-well microtitration plates: an aide to rapid identification of transformed cell clones. 96孔微滴定板中激素刺激腺苷酸环化酶的原位测定:快速鉴定转化细胞克隆的辅助手段。

Journal of receptor research Pub Date : 1993-01-01 DOI: 10.3109/10799899309073646

A P Themmen, V Hinrichs, M Birnbaumer

{"title":"In situ assay of hormone-stimulated adenylyl cyclase in 96-well microtitration plates: an aide to rapid identification of transformed cell clones.","authors":"A P Themmen, V Hinrichs, M Birnbaumer","doi":"10.3109/10799899309073646","DOIUrl":"https://doi.org/10.3109/10799899309073646","url":null,"abstract":"An in situ assay able to detect hormonally stimulated or inhibited adenylyl cyclase (AC) activity on as few as 5,000 cells/well has been developed. In addition the assay monitors phosphatase activity which serves as a marker for cell density. Cells are plated in replicate wells at least one day before the assay, and the medium containing AC reagents, an ATP regenerating system, a PDE inhibitor, additives that regulate receptors and/or G proteins, and 5 mM p-nitrophenyl phosphate (pNPP), plus Tris buffer to pH 7.5 is added. The hypotonic medium causes permeabilization of cells without massive lysis. After stopping the reaction with 100 microliters of a solution with ATP, cAMP and SDS, the color indicating phosphatase activity is quantified by an ELISA reader, and AC activity measured by standard methods. At proper cell density (pNPP hydrolysis) the assay shows proportionality up to 2 hours. The assay is particularly useful in transfection experiments. As few as 50,000 cells can be plated and identified as receptor \"positive\" or receptor \"negative\". The assay was key to our cloning of the V2 AVP receptor. The assay accelerated the preparation of stable cell lines with LH, FSH, adrenergic and serotonin 1D beta/1B and 1E receptors. It should also be useful in studies in which the transfected cDNA encodes the adenylyl cyclase proper.","PeriodicalId":16948,"journal":{"name":"Journal of receptor research","volume":"13 1-4","pages":"69-78"},"PeriodicalIF":0.0,"publicationDate":"1993-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.3109/10799899309073646","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"19369027","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 6

A partially automated radioligand binding assay system for use in clinical and pharmaceutical research. 用于临床和药物研究的部分自动化放射配体结合测定系统。

Journal of receptor research Pub Date : 1993-01-01 DOI: 10.3109/10799899309073667

B Liebl, T Anhäupl, E Haen, B Günster, M Georgieff

{"title":"A partially automated radioligand binding assay system for use in clinical and pharmaceutical research.","authors":"B Liebl, T Anhäupl, E Haen, B Günster, M Georgieff","doi":"10.3109/10799899309073667","DOIUrl":"https://doi.org/10.3109/10799899309073667","url":null,"abstract":"Using a Tecan robotic sample processor and IBM compatible PCs we have developed a flexible, partially automated radioligand binding assay system. It handles pipetting parameters of up to 16 saturation or competition experiments at a time with up to 24 radioligand- or competitor-concentrations in a range over 4 orders of magnitude per experiment. The system provides enough flexibility so that all pipetting parameters including different tube-, rack-sizes, sample volumina and pipetting sequences may be easily adapted to the large variety of experimental requirements in binding assays. It rationalizes and increases assay throughput (up to 70% spare of working time), improves reliance and reproducibility of results. Radioactive exposure is minimized to the time preparing the radioligand working solution and transferring the sample tubes to and from the sample processor. The system has proven effective in various investigations on binding interactions, as well as in clinical studies on receptor expression under physiologic, pathological and therapeutic conditions.","PeriodicalId":16948,"journal":{"name":"Journal of receptor research","volume":"13 1-4","pages":"369-78"},"PeriodicalIF":0.0,"publicationDate":"1993-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.3109/10799899309073667","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"19369686","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 5

Calcium induced modulation of peripheral-type benzodiazepine receptors in rat kidney membranes. 钙诱导大鼠肾膜外周型苯二氮卓受体的调节。

Journal of receptor research Pub Date : 1993-01-01 DOI: 10.3109/10799899309073704

D J Calvo, J H Medina

引用次数: 0

Chronic antagonist treatment does not alter the mode of interaction of dopamine with rat striatal dopamine receptors. 慢性拮抗剂治疗不改变多巴胺与大鼠纹状体多巴胺受体的相互作用模式。

Journal of receptor research Pub Date : 1993-01-01 DOI: 10.3109/10799899309073664

K M O'Boyle, K T Gavin, N Harrison

引用次数: 8

Structural specificity of polyamines in modulating the binding of estrogen receptor to potential Z-DNA forming sequences. 多胺调节雌激素受体与潜在Z-DNA形成序列结合的结构特异性。

Journal of receptor research Pub Date : 1993-01-01 DOI: 10.3109/10799899309063267

T Thomas, T J Thomas

{"title":"Structural specificity of polyamines in modulating the binding of estrogen receptor to potential Z-DNA forming sequences.","authors":"T Thomas, T J Thomas","doi":"10.3109/10799899309063267","DOIUrl":"https://doi.org/10.3109/10799899309063267","url":null,"abstract":"Estrogen receptor (ER) is a gene-regulatory protein that mediates the action of estradiol. In order to examine the role of conformational dynamics of DNA in estrogenic regulation of gene expression, we studied the binding of ER to poly(dA-dC).poly(dG-dT) which undergoes transition to a left-handed Z-DNA form. This type of dinucleotide repeats are widely distributed in mammalian genome and are present in estrogen response elements. Binding affinity of ER for the polynucleotide was assessed by its ability to release ER bound to DNA-cellulose. ER binding by poly(dA-dC).poly(dG-dT) was enhanced in the presence of an endogenous polyamine, spermidine, H2N(CH2)4NH(CH2)3NH2. The concentration of spermidine required for facilitating 50% elution of ER (EC50) was 75 microM. This EC50 increased to 500 microM for a spermidine homolog, H2N(CH2)8NH(CH2)3NH2, demonstrating polyamine structural specificity. Spectroscopic measurements showed that the presence of 100-200 microM spermidine initiated changes in the conformation of the polynucleotide indicative of Z-DNA form, but a major alteration to Z-DNA spectrum occurred only at 300 microM concentration. These data suggest that ER favors DNA sequences poised for Z-DNA transition. The efficacy of spermidine homologs in facilitating ER-DNA interaction may be important in predicting their efficiency to replace cellular functions of spermidine.","PeriodicalId":16948,"journal":{"name":"Journal of receptor research","volume":"13 8","pages":"1115-33"},"PeriodicalIF":0.0,"publicationDate":"1993-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.3109/10799899309063267","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"19242498","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 16

Towards an identification of odorant receptors. 气味受体的识别。

Journal of receptor research Pub Date : 1993-01-01 DOI: 10.3109/10799899309073677

H Breer, K Raming, J Krieger, I Boekhoff, J Strotmann

引用次数: 3

Vasopressin receptors in adrenal cortex of sheep: does autoradiography indicate an irreversible binding of the ligand? 绵羊肾上腺皮质的抗利尿激素受体:放射自显影显示配体的不可逆结合吗?

Journal of receptor research Pub Date : 1993-01-01 DOI: 10.3109/10799899309073660

R A Lutz, G Tomasz, S Lüem, P Blum, V Pliska

{"title":"Vasopressin receptors in adrenal cortex of sheep: does autoradiography indicate an irreversible binding of the ligand?","authors":"R A Lutz, G Tomasz, S Lüem, P Blum, V Pliska","doi":"10.3109/10799899309073660","DOIUrl":"https://doi.org/10.3109/10799899309073660","url":null,"abstract":"Tritiated arginine vasopressin ([3H]-AVP) labelled specific loci of murine renal medulla and ovine adrenal cortex in thin sections of an autoradiographic experiment. The label was fully displaced by 2 x 10(-6) M cold ligand in the case of renal, but not of adrenal sections. 10 and 100 microM AVP, however, partially displaced the radioactivity also from labelled adrenal sections. At room temperature, the half maximal blackening of the film occurred at a concentration of 26 +/- 0.9 microM. In binding experiments employing AVP and adrenocortical cell membranes, the model assuming two saturable binding sites yielded a significantly better fit than the one-site model. The equilibrium dissociation constants of ice-cold membrane preparations were 8.67 nmol/l for the high affinity site and 3.16 mumol/l for the low affinity binding site. It is concluded that the low affinity binding is governed by laws of chemical equilibrium, rather than by surface adsorption or similar \"nonspecific\" phenomena. When such low affinity sites are present in a tissue, higher concentrations of cold ligand ought to be used before a nondisplaceable binding is ascribed as \"non-specific\" or \"irreversible\".","PeriodicalId":16948,"journal":{"name":"Journal of receptor research","volume":"13 1-4","pages":"283-93"},"PeriodicalIF":0.0,"publicationDate":"1993-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.3109/10799899309073660","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"19434374","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 1

Selective down-regulation of the agonist-dependent high affinity state of rat cortical 5-HT2 receptors following chronic treatment with amperozide. 慢性安哌唑治疗后大鼠皮质5-HT2受体激动剂依赖性高亲和力状态的选择性下调。

Journal of receptor research Pub Date : 1993-01-01 DOI: 10.3109/10799899309073668

J O Svartengren

{"title":"Selective down-regulation of the agonist-dependent high affinity state of rat cortical 5-HT2 receptors following chronic treatment with amperozide.","authors":"J O Svartengren","doi":"10.3109/10799899309073668","DOIUrl":"https://doi.org/10.3109/10799899309073668","url":null,"abstract":"Amperozide, a putative antipsychotic drug, is a selective serotonin 5-HT2 receptor antagonist with moderate affinity for the rat brain dopamine D2 receptor. The aim of the present study was to investigate the regulation of 3H-ketanserin binding of the rat cerebral cortex 5-HT2 receptor following chronic amperozide treatment. Amperozide (5 mg/kg/day p.o. for three weeks) significantly reduced the number of 5-HT2 receptors in rat cerebral cortex (172.9 +/- 8.3 vs 128.4 +/- 4.8 fmol/mg protein; p < 0.001, n = 5-6). There was no change of Kd of 3H-ketanserin. The agonist serotonin (5-HT) recognized two sites of 3H-ketanserin binding in membranes from both saline- and amperozide-treated rats as analysed by non-linear regression using LIGAND. The concentration of high affinity sites was reduced by 51% (53.1 +/- 2.9 vs 25.7 +/- 3.7 fmol/mg protein; p < 0.01, n = 3) while the amount of the low affinity sites remained unchanged. Serotonin displacement of 3H-ketanserin binding was sensitive to regulation by GTP. The present data indicated that GTP regulation was partially lost following chronic amperozide treatment. Displacement of the antagonists methysergide and amperozide were not affected by GTP. In conclusion, the down-regulation of the 5-HT2 receptors appears mainly due to a decrease of the serotonin-recognized high affinity state, and the loss of GTP sensitivity may indicate a decreased ability to form the high affinity state of the receptor. Such a molecular regulation might underlie an altered 5-HT responsiveness and thus be of therapeutic value.","PeriodicalId":16948,"journal":{"name":"Journal of receptor research","volume":"13 1-4","pages":"379-92"},"PeriodicalIF":0.0,"publicationDate":"1993-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.3109/10799899309073668","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"19434377","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 13

A novel agonist binding site on nicotinic acetylcholine receptors. 烟碱乙酰胆碱受体上一种新的激动剂结合位点。

Journal of receptor research Pub Date : 1993-01-01 DOI: 10.3109/10799899309073670

E F Pereira, M Alkondon, T Tano, N G Castro, M M Fróes-Ferrão, R Rozental, R S Aronstam, A Schrattenholz, A Maelicke, E X Albuquerque

{"title":"A novel agonist binding site on nicotinic acetylcholine receptors.","authors":"E F Pereira, M Alkondon, T Tano, N G Castro, M M Fróes-Ferrão, R Rozental, R S Aronstam, A Schrattenholz, A Maelicke, E X Albuquerque","doi":"10.3109/10799899309073670","DOIUrl":"https://doi.org/10.3109/10799899309073670","url":null,"abstract":"This report provides evidence that physostigmine (Phy) and benzoquinonium (BZQ) are able to activate nicotinic acetylcholine receptors (nAChRs) through binding site(s) distinct from those of the natural transmitter, ACh. Such findings are in agreement with a second pathway of activation of nAChRs. Receptor activation may be modulated through the novel site, and, consequently, physiological processes involving nicotinic synapses could be controlled. Using patch clamp techniques, single channel currents activated by ACh and anatoxin were recorded from frog interosseal muscle fibers under cell-attached condition and outside-out patches excised from cultured rat hippocampal neurons. Whole cell nicotinic currents were also studied in the cultured neurons. In most of the neurons, nicotinic responses were blocked by the nicotinic antagonists methyllycaconitine (MLA) and alpha-bungarotoxin (alpha-BGT). Evaluation of the effects of Phy and BZQ on the muscle and on the alpha-BGT- and MLA-sensitive neuronal nAChRs demonstrated that both compounds were open channel blockers at these receptors. Furthermore, at low micromolar concentrations, Phy and BZQ activated the nAChRs of all preparations tested, such an effect being unexpectedly resistant to alpha-BGT or MLA. Thus, the nAChRs could be activated via two distinct binding sites: one for ACh and the other for Phy and BZQ. These findings and previous biochemical results led us to suggest that a putative endogenous ligand could bind to the new site and thereby regulate the activation of nAChRs in nicotinic synapses.","PeriodicalId":16948,"journal":{"name":"Journal of receptor research","volume":"13 1-4","pages":"413-36"},"PeriodicalIF":0.0,"publicationDate":"1993-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.3109/10799899309073670","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"19434378","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 48

Comparison of the binding and functional actions of angiotensin agonists in clone 9 cells: additional evidence for angiotensin II receptor heterogeneity. 血管紧张素激动剂在克隆细胞中的结合和功能作用的比较:血管紧张素II受体异质性的额外证据。

Journal of receptor research Pub Date : 1993-01-01 DOI: 10.3109/10799899309063262

M R Kozlowski, M Arcuri, L Zynardi

{"title":"Comparison of the binding and functional actions of angiotensin agonists in clone 9 cells: additional evidence for angiotensin II receptor heterogeneity.","authors":"M R Kozlowski, M Arcuri, L Zynardi","doi":"10.3109/10799899309063262","DOIUrl":"https://doi.org/10.3109/10799899309063262","url":null,"abstract":"The effects of the angiotensin-II (AII) agonists and antagonists on both 125I-SARILE binding and phosphoinositol (PI) accumulation in clone 9 cells were examined. Clone 9 cells, which are derived from rat liver, have been shown to respond to AII agonists with an increase in PI accumulation which is inhibitable by Sar1,Ile8-AII (SARILE) and DUP-753 but not PD-123319, suggesting that they possess the AT1 subtype of AII receptor. The present results confirmed these properties. The order of potency of AII agonists was AII > AIII > AI. Clone 9 cells also possessed binding sites for 125I-SARILE. The majority of these were AT1 type receptors, although a small number of AT2 receptors may also have been present. The order of potency of AII agonists in inhibiting 125I-SARILE binding was AII >> AIII = AI. The difference in rank order of potency between the functional and binding assay was due to AIII being much less potent in the binding assay than the functional assay. Since the potency of AIII relative to AII was lower than that at either AT1 or AT2 subtypes of AII receptor, these data suggest that an additional subtype, with selectively low affinity for AIII, exists.","PeriodicalId":16948,"journal":{"name":"Journal of receptor research","volume":"13 7","pages":"1031-40"},"PeriodicalIF":0.0,"publicationDate":"1993-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.3109/10799899309063262","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"19380538","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 3