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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Acta neuropathologica 82 (1991), S. 516-519 
    ISSN: 1432-0533
    Keywords: Multidrug resistance ; P-glycoprotein ; Glial tumor ; Immunohistochemistry ; RNA analysis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary The most consistantly reported alteration of multidrug-resistant carcinoma cells is the overexpression of a membrane glycoprotein, termed P-glycoprotein. In this study we examined whether the strong intrinsic chemotherapy resistance of glial tumors might be related to the expression of the MDR1 gene which codes for P-glycoprotein. Fourteen glial tumors were examined immunohistochemically using the monoclonal antibody C219. In addition, RNA samples of 11 of these tumors were analysed using a sensitive Northern blot assay. P-glycoprotein is expressed in all 14 glial tumors; the number of stained tumor cells, however, varied considerably ranging from 0.3% to 15%. There was no correlation between the number of MDR1-positive cells and the histological malignancy. Varying amounts of MDR1 mRNA were detectable in 7 from 11 examined tumors. The results of our study show that the MDR1 gene is expressed in human glial tumors and suggest that the multidrug transporter may contribute to the clinical non-responsiveness of these tumors to chemotherapy.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    Experimental brain research 69 (1987), S. 208-212 
    ISSN: 1432-1106
    Keywords: Prenatal development ; Messenger RNA ; Brain ; Opioids ; Pig
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Using solution hybridization analysis and Northern blotting with complementary RNA probes labelled to high specific activity, levels of proenkephalin A and B mRNA were analyzed throughout prenatal development in the hippocampus and striatum of fetal pigs. A differential time course for the appearance of these opioid precursor mRNAs was observed: in hippocampus, both mRNAs increased linearly throughout development with proenkephalin B mRNA increasing faster than proenkephalin A mRNA. In striatum, both mRNAs behaved similarly, increasing to a maximum level around mid-gestation and declining thereafter. The differences might be attributed to differential localization of the two precursor systems in the tissues and might be of functional relevance.
    Type of Medium: Electronic Resource
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  • 3
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Science Ltd
    Journal of neurochemistry 66 (1996), S. 0 
    ISSN: 1471-4159
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Abstract: The actions of exogenous and endogenous opioids are mediated by at least three different opioid receptors, called μ, κ, and δ. Recently, we have detected a new variant of the rat μ-opioid receptor, which we termed rMOR1B and which differs from rMOR1 (now also called rMOR1A) in the amino acid sequence at the C-terminus. Both isoforms were proposed to be splicing variants of the same gene. To elucidate the molecular mechanism leading to the formation of the new variant, the exon/intron structure of the rat μ-opioid receptor gene in the respective area has been determined by analyzing a genomic P1 phage clone. In addition, we have investigated the putative promoter region of this gene. The present study revealed that rMOR1B is generated by an alternative splicing event whereby a previously unknown exon will be placed behind exon 3 to form rMOR1B mRNA, which is separated from the latter by an intron. Therefore, this new exon has to be called exon 4, whereas the former exon 4, which encodes the C-terminus of MOR1A, now becomes exon 5. Examination of the putative rat promoter region revealed a high degree of nucleotide sequence homology to the mouse gene. Using an RNase protection approach, one single transcription initiation site could be located at 230 bp upstream of the translation start. This is similar to the situation in the mouse, where four major transcription start sites were reported to lie close together around 270 bp upstream of the protein coding region.
    Type of Medium: Electronic Resource
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  • 4
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    Annals of the New York Academy of Sciences 467 (1986), S. 0 
    ISSN: 1749-6632
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Natural Sciences in General
    Type of Medium: Electronic Resource
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  • 5
    Electronic Resource
    Electronic Resource
    Springer
    Journal of molecular medicine 70 (1992), S. 224-231 
    ISSN: 1432-1440
    Keywords: Nicotine ; Pro-opiomelanocortin ; Proenkephalin ; Prodynorphin ; Vasopressin ; α3 nicotinic receptor ; Gene expression ; AdAenal ; Pituitary ; Hypothalamus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary The effect of acute and chronic nicotine treatment of rats on the mRNA levels coding for the three opioid peptide precursors, for provasopressin and for the a3 subunit of nicotinic receptors in brain, pituitary and/or adrenal medulla of rats was investigated. Nicotine was found to increase the levels of proenkephalin mRNA in the adrenal medulla, but did not affect the levels of PENK mRNA in striatum, hypothalamus and hippocampus. The mRNA levels of prodynorphin were increased together with that of provasopressin in the hypothalamus after nicotine, whereas the prodynorphin mRNA levels in the hippocampus and the striatum remained unchanged. Nicotine treatment resulted in an increase in the pro-opiomelanocortin mRNA levels in the anterior pituitary and in a decrease in the intermediate pituitary, but did not change the levels of pro-opiomelanocortin mRNA in the hypothalamus. The levels of mRNA coding for the α3 subunit of nicotinic receptors in the hypothalamus and the adrenal medulla remained unchanged. The increase in the prodynorphin and provasopressin mRNA levels in the hypothalamus was most pronounced 1 day after s.c. application of two doses of 0.4 mg/kg nicotine (about 100% above control). A smaller increase in mRNA concentrations (about 30%) was found after tonic infusion of the drug for 4 days (4 mg/kg per day), whereas no change was observed after tonic infusion of nicotine for 7 and 14 days indicating the development of complete tolerance. The increase in proenkephalin mRNA levels in the adrenal medulla was highest after the short-term application of nicotine (about 150% above control). Less, but still significant increases in the mRNA levels (about 40%) were also seen after 7 and 14 days of tonic nicotine administration suggesting that no complete desensitization is developing to the effect of nicotine. This desensitization appeared to be less pronounced when the drug was applied in a pulsatile manner using minipumps, since a high increase in the PENK mRNA levels (100% above control levels) was observed after intermittent infusion of nicotine (six boli per day of 1 mg/kg for 7 days). These findings demonstrate that nicotine can alter gene expression of opioid peptides and vasopressin in certain rat tissues, and that the mode of drug administration plays an important role in this effect of nicotine. The possibility that the ‘pulsatile’ administration of nicotine by cigarette smokers may also result in an altered expression of opioid peptide genes in humans is discussed.
    Type of Medium: Electronic Resource
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  • 6
    Electronic Resource
    Electronic Resource
    Springer
    Journal of molecular medicine 72 (1994), S. 925-929 
    ISSN: 1432-1440
    Keywords: Nicotine ; Proenkephalin ; c-fos ; Gene expression ; Chromaffin cells
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The induction of the proenkephalin gene by nicotine has been characterized in bovine adrenal medullary chromaffin cells. Nicotine (10 μM) caused an approximately fourfold increase in the proenkephalin mRNA levels within 24 h. The half-life of the proenkephalin mRNA in nicotine-stimulated cells was similar to that in control cells (about 13 h), indicating that nicotine does not affect mRNA stability but acts at the levels of proenkephalin gene transcription. This was also supported by experiments showing that the expression of a proenkephalin chloramphenicol acetyl transferase reporter gene (PENKCAT-153/+50) containing 153 nucleotides of upstream promoter sequences is increased (about twofold) by nicotine after transient transfection in the chromaffin cells. In addition, nicotine induced a marked elevation of the immediate early gene mRNAs c-fos, c-jun, and jun-B. Maximally increased levels for c-fos mRNA (about 100-fold) were obtained after 20 min. c-jun and jun-B were increased three- to fivefold 60 min after nicotine addition. The expression of PENKCAT-153/+53 and of a proenkephalin gene reporter plasmid which contains a dimer of the enkephalin cAMP responsive element 2 (ENKCRE-2) in front of a minimal promoter was increased by cotransfection of a c-fos expression plasmid, indicating that nicotine may induce the proenkephalin gene in chromaffin cells via c-Fos which binds to the ENKCRE-2 element.
    Type of Medium: Electronic Resource
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  • 7
    ISSN: 1432-1912
    Keywords: Vasopressin ; Oxytocin ; Opioid peptides ; In vitro release ; Pituitary
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Dynorphin (1–17), and to a lesser extent, β-endorphin and [Leu]enkephalin (10−6 M each) decreased the spontaneous release of vasopressin (VP) from the rat neurointermediate pituitary in vitro, whereas the oxytocin (OT) release remained unchanged. Naloxone, however, did not significantly alter the spontaneous VP and OT release. Dynorphin (1–17) (10−7 M) increased the electrically evoked release of VP and OT, while 10−6 M had a significant, somewhat less pronounced stimulatory effect only on VP, but not on OT release. The opiate inactive fragment [des-Tyr1]dynorphin (1–17) did not change the evoked VP and OT release, indicating that the dynorphin effect was mediated by opiate receptors. β-Endorphin (10−6 M and 10−7 M) did not alter the evoked VP and OT secretion. 10−6 M [Leu]enkephalin induced a stimulation of the evoked OT, but not VP release; 10−7 M [Leu]enkephalin had no effect, neither on VP nor on OT release. The opiate antagonist naloxone 10−5 M) induced an increase in the evoked VP and, even more pronounced, OT release. In a concentration of 10−6 M, however, naloxone only increased the evoked OT release. When naloxone and dynorphin (1–17) were concomitantly applied, their stimulatory effects on the evoked VP and OT release were additive. Similarly to the effects of naloxone, addition of a monoclonal antibody which binds to the common N-terminal sequence of all endogenous opioid peptides, resulted in a marked increase in the evoked secretion of VP and, to an even more pronounced degree, of OT. Thus, the stimulated VP and OT release seems to be under a tonic inhibitory control of opioids. Electrical stimulation was found to enhance the release of endogenous immunoreactive dynorphin from the neurointermediate pituitary, while the release of immunoreactive β-endorphin was even decreased. In conclusion, the present data indicate a dual role of endogenous opioid peptides in the VP and OT secretion at the level of the posterior pituitary.
    Type of Medium: Electronic Resource
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  • 8
    Electronic Resource
    Electronic Resource
    Springer
    Naunyn-Schmiedeberg's archives of pharmacology 288 (1975), S. 163-177 
    ISSN: 1432-1912
    Keywords: Protein Binding ; Opiates ; Hydrophobic Interaction
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary The unspecific binding of four narcotic analgesics 3H-dihydromorphine, 14C-morphine, 3H-etorphine and 3H-fentanyl to human albumin, human plasma, rabbit plasma and several tissue homogenates from rabbits was investigated using equilibrium dialysis and ultrafiltration. At a drug concentration of 10−7 M in human plasma, dihydromorphine is bound to an extent of 14%, morphine to 23%, etorphine to 88% and fentanyl to 70%. These differences in binding are due to different degrees of hydrophobic interaction between the drugs investigated and the plasma or tissue components. The hydrophobic interactions are due to the unionized form of the drugs. The ionized form is bound to a negligible extent with all four compounds, possibly in part by ionic mechanism. Binding increased with increasing ionic strength of the protein solution, with raising temperature between 0°C and 37°C and with increasing pH values of the protein solution, features which are characteristic of hydrophobic interactions. Scatchard plots of the binding data, from which the total binding constants nk were derived, indicated high concentrations of binding sites compared with drug concentrations found analgesically effective in vivo.
    Type of Medium: Electronic Resource
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  • 9
    ISSN: 1432-2072
    Keywords: Precipitated morphine withdrawal ; Morphine antagonists and partial agonists ; Stereospecific opiate binding ; Rats
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract In morphine-dependent rats the withdrawal precipitating properties of various morphine antagonists and partial agonists were studied by quantitatively evaluating a variety of different withdrawal signs. A comparison of the dose response curves of the various substances obtained for the different signs revealed marked differences in respect to the lowest effective doses (EDs) necessary to precipitate the withdrawal signs as well as in the maximum frequencies of the signs induced. The “pure” antagonist, naloxone, which was judged very potent according to the ED, precipitated the lowest levels of jumping, whereas certain partial agonists of the benzomorphane type, which were less potent according to the ED, induced very high levels of this sign. These latter compounds, however, failed to precipitate “complete” withdrawal, as evidenced by the nearly complete absence of some of the withdrawal signs. The jumping precipitating potency of the antagonists as judged from the ED was found to be highly correlated to the stereospecific binding of these substances to rat brain homogenate. On the other hand, the ability of the substances to precipitate high levels of jumping was seen to increase, at least within a certain range, with increasing degree of agonistic properties, as indicated by the ratio of stereospecific binding in the presence and absence of sodium.
    Type of Medium: Electronic Resource
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  • 10
    ISSN: 1432-2072
    Keywords: Key words Hippocampus ; Lesion ; Glutamate receptor and release ; Dopamine receptor ; Locomotor activity ; Schizophrenia ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract In 6-week and 8-week-old rats (pre- and postpubertally) with neonatal excitotoxic lesions of the ventral hippocampus with ibotenic acid (IBO), we have studied apomorphine-induced motor activity and glutamate and dopamine D1 and D2 binding sites in the hippocampus, striatum, nc. accumbens and frontal cortex as well as K+-stimulated (3H)-D-aspartate release from hippocampal and frontal cortical slices. Specific glutamate binding was enhanced in the frontal cortex of 8-week-old IBO-treated animals, whereas that in other brain regions remained unchanged. Both D1 and D2 binding sites were downregulated in the striatum without changes in other brain structures. In 6-week-old rats, neither the glutamate nor the dopamine binding sites were altered. The amino acid release from hippocampal and frontal cortical slices of adult IBO treated rats was significantly decreased in comparison to controls, whereas in 6-week-old rats, no significant alterations were detectable. The additionally monitored motor activity was enhanced only in adult IBO-lesioned rats after apomorphine pretreatment. The present data are in agreement with the hypothesis of hyperactive dopamine and hypoactive glutamate systems in schizophrenia and are discussed in the light of schizophrenia-like behavioral changes in rats after postnatal hippocampal IBO lesion.
    Type of Medium: Electronic Resource
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