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  • 1
    Electronic Resource
    Electronic Resource
    [s.l.] : Nature Publishing Group
    Nature 292 (1981), S. 623-625 
    ISSN: 1476-4687
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Chemistry and Pharmacology , Medicine , Natural Sciences in General , Physics
    Notes: [Auszug] Uteri from immature New Zealand rabbits (typically six rabbits per experiment) were pooled, the endometrium removed and myometrial strips divided equally into two groups. Myometrium in the first group was used immediately in isometric contraction studies or was homogenized and a particulate ...
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    Molecular and cellular biochemistry 67 (1985), S. 47-53 
    ISSN: 1573-4919
    Keywords: α1-adrenergic receptor ; membrane ; photoaffinity ; smooth muscle
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Summary In this study, we have used an α1-adrenergic receptor photoaffinity ligand, 2-[4-(4-azido-3-iodo-benzoyl)-piperazin-1-yl]-4-amino-6, 7-dimethoxyquinazoline (125I-APD), to label covalently the α1-adrenergic receptor in a smooth muscle cell line. Our results indicate that in the absence of light, (125I)APD binds reversibly to a site in the DDT1 MF-2 cell membranes having pharmacological characteristics of an α1-adrenergic receptor. Following incorporation of (125I)ADP into partially purified membranes a single labeled band of protein with a Mr of 81 000 was visualized by autoradiography following sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Incorporation of (125I)-APD into this band was affected by adrenergic agonists and antagonists in a manner consistent with an α1-adrenergic interaction. Prazosin (α1-selective) blocked incorporation of the label into the Mr = 81 000 protein while yohimbine (α2-selective) did not. Of the adrenergic agonists, (−)-epinephrine and (−)-norepinephrine but not (−)-isoproterenol blocked labeling of the Mr − 81 000 protein. We conclude that the ligand binding site of the DDT1 MF-2 cell α1-adrenergic receptor resides in a Mr = 81 000 protein.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1573-4919
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract We have demonstrated that glucocorticoids induce in DDT1 MF-2 cells by a glucocorticoid mediated mechanism the synthesis of a methionine-cysteine rich protein of 29 000 Mr (p29). Induction of p29 is not observed in DDT1 MF-2 GR glucocorticoid resistant variants which have only 7% of glucocorticoid receptor site per cell compared to wild type cells. Increased synthesis of p29 is specific to glucocorticoids since neither androgens, estrogens, progesterone nor the glucocorticoid antagonist dexamethasone mesylate are effective inducers. Stimulation of p29 synthesis in wild type cells is observed at 10−10 M triamcinolone acetonide, reaching a maximum at a concentration of 1 × 10−8 M. The induction of p29 is not a function of glucocorticoid arrest of DDT1 MF-2 cells since DDT1 MF-2 cells promoted to re-enter the cell cycle by 50 ng/ml platelet derived growth factor (PDGF) continue synthesis of p29. Finally, increased levels of p29 translation products are observed in cell free translation assays carried out utilizing poly A− RNA transcripts isolated from glucocorticoid treated cells. These data suggest that the glucocorticoid stimulation of p29 synthesis is a transcriptional and/or RNA processing event controlled by glucocorticoid receptor complexes.
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1573-4919
    Keywords: glucocorticoid receptor ; β-adrenergic receptor regulation ; smooth muscle ; adenylate cyclase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract We have shown that glucocorticoids induce the appearance of β2-adrenergic receptors in membranes of the ductus deferens smooth muscle cell line (DDT1 MF-2). A concomitant increase in isoproterenol stimulated adenylate cyclase activity in the absence of exogenously applied GTP was observed as was a significantly increased (p 〈 0.05) sensitivity of the adenylate cyclase system to exogenously applied GTP. However, no significant difference in the maximal velocity of adenylate cyclase between control and steroid treatment was measurable in the presence of sodium fluoride. Induction of β2-adrenergic receptors in DDT1 MF-2 cells is correlated with the presence of steroid receptors (androgen and glucocorticoid) in the cells since estrogens and progesterones had no effect on receptor levels. Finally, utilizing dense amino acid labeling of cells to measure old versus newly synthesized receptor sites by a density shift method, we have documented that glucocorticoid induction of β2-adrenergic receptors involves synthesis of new receptor protein.
    Type of Medium: Electronic Resource
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  • 5
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: In the liver, transcript destabilization contributes to the decrease in steady-state levels of β2-adrenergic receptor mRNA that occurs during early postnatal development in the rat. From genomic DNA, polymerase chain reaction (PCR) was used to amplify a 718-basepair (bp) fragment of the β2-adrenergic receptor gene including the entire 3′-untranslated region. Results from SDS-gel electrophoresis and autoradiography demonstrated a Mr 85,000 cellular factor present in postnatal day 60, but not fetal day 18 rat liver that was ultraviolet (UV) light-crosslinked to in vitro transcribed β2-adrenergic receptor RNA 3′-untranslated region. Unlabeled β2-adrenergic receptor RNA 3′-untranslated region, but not mouse β-actin RNA, competed with labeled β2-adrenergic receptor RNA 3′-untranslated region for binding to the Mr 85,000 protein. Cross-linking of the β2-adrenergic receptor RNA 3′-untranslated region to the Mr 85,000 protein was inhibited by the ribohomopolymer poly(U), with poly(A), poly(C) and poly(G) having little or no effect. Thus, a Mr 85,000 protein has been identified in adult male rat liver that may interact with U-rich sequences in the 3′-untranslated region of the β2-adrenergic receptor mRNA and may account for the decreased stability of hepatic β2-adrenergic receptor gene transcripts that occurs during development. © 1995 Wiley-Liss, Inc.
    Additional Material: 5 Ill.
    Type of Medium: Electronic Resource
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  • 6
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 157 (1993), S. 70-76 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Hepatic responsiveness to β2-adrenergic stimulation is dynamically regulated during early development as well as following hepatic injury and disease. In the present study, the molecular mechanisms that underlie the decline in the steady-state levels of hepatic β2-adrenergic receptor mRNA that occurs during development in the male rat were investigated. As determined by nuclear run-on assays, an age-associated reduction in β2-adrenergic receptor gene transcription was observed. The transcription rate of the β2-adrenergic receptor gene in postnatal day 18 liver was approximately 50% lower than that of fetal liver. Stability of β2-adrenergic receptor gene transcripts was highest (t1/2 ≈ 6 h) in hepatocytes isolated from fetal rats and was lowest (t1/2 ≈ 1 h) in hepatocytes isolated from postnatal day 14 rats. In fetal hepatocytes, but not postnatal day 2 hepatocytes, cycloheximide appeared to stabilize β2-adrenergic receptor gene transcripts in the presence of actinomycin D. These findings establish the molecular basis of reduced steady-state levels of β2-adrenergic receptor mRNA in liver during early postnatal development and suggest multilevel regulatory control of hepatic β2-adrenergic receptor gene expression. © 1993 Wiley-Liss, Inc.
    Additional Material: 4 Ill.
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  • 7
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 147 (1991), S. 55-61 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Metabolic events stimulated by epinephrine and norepinephrine in hepatocytes isolated from fetal and early postnatal male rats are largely mediated through the β2-adrenergic receptor-/cyclic AMP dependent-system, whereas the same stimuli are transduced through the α1-adrenergic receptor-/phosphatidylinositol dependent-system in hepatocytes isolated from young adult male rats. This developmental transition was investigated by correlating hepatic α1- and β2-adrenergic receptor gene transcript levels with receptor levels as determined with selective radioligands in livers from late fetal to postnatal day 120 male Sprague-Dawley rats. β2-Adrenergic receptor concentration, initially high in membrane preparations isolated from fetal livers (203 ± 21 fmol/mg protein), dropped precipitously n postnatal day 6 livers (14± 2 fmol/mg protein) and remained low throughout development out to postnatal day 90.β-Adrenergic receptor mRNA levels were highest in fetal livers, were decreased somewhat in postnatal day 6 livers and were uncetectable in livers beyond postnatal day 15. In contrast, hepatic α-adrenergic receptor concentration was relatively low in fetal livers (86 ± 25 fmol/mg protein) and remained low until postnatal day 18. Thereafter, a steady increase in α1-adrenergic receptors was observed until adult levels. (270 ± 24 fmol/mg protein) were achieved at postnatal day 27. α1-Adrenergic receptor mRNA levels increased ∼ 3-fold, reaching a peak at postnatal day 24. Surprisingly, at postnatal day 30 hepatic α1-adrenergic receptor mRNA levels dropped to fetal levels; but, gradually increased with continued development. Thus, hepatic α1- and β2-adrenergic receptors appear to be under complex regulatory control which may include transcriptional, as well as post-transcriptional, mechanisms.
    Additional Material: 4 Ill.
    Type of Medium: Electronic Resource
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