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See detailDisrupting the melanin-concentrating hormone receptor 1 in mice leads to cognitive and NMDA response deficit
Grisar, Thierry ULg; Adamantidis, Antoine ULg; Thomas, Elizabeth et al

in Journal of the Neurological Sciences (2005, November 15), 238(Suppl. 1), 288

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See detailPig tissues express a catalytically inefficient 25-kDa thiamine triphosphatase: Insight in the catalytic mechanisms of this enzyme
Szyniarowski, Piotr; Lakaye, Bernard ULg; Czerniecki, Jan ULg et al

in Biochimica et Biophysica Acta - General Subjects (2005), 1725(1), 93-102

Thiamine triphosphate (ThTP) is found in most organisms and may be an intracellular signal molecule produced in response to stress. We have recently cloned the cDNA coding for a highly specific mammalian ... [more ▼]

Thiamine triphosphate (ThTP) is found in most organisms and may be an intracellular signal molecule produced in response to stress. We have recently cloned the cDNA coding for a highly specific mammalian 25-kDa thiamine triphosphatase. The enzyme was active in all mammalian species studied except pig, although the corresponding mRNA was present. In order to determine whether the very low ThTPase activity in pig tissues is due to the absence of the protein or to a lack of catalytic efficiency, we expressed human and pig ThTPase in E. coli as GST fusion proteins. The purified recombinant pig GST-ThTPase was found to be 2-3 orders of magnitude less active than human GST-ThTPase. Using site-directed mutagenesis, we show that, in particular, the change of Glu85 to lysine is responsible for decreased solubility and catalytic activity of the pig enzyme. Immunohistochemical studies revealed a distribution of the protein in pig brain very similar to the one reported in rodent brain. Thus, our results suggest that a 25-kDa protein homologous to hThTPase but practically devoid of enzyme activity is expressed in pig tissues. This raises the possibility that this protein may play a physiological role other than ThTP hydrolysis. [less ▲]

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See detailDisrupting the melanin-concentrating hormone receptor 1 in mice leads to cognitive deficits and alterations of NMDA receptor function.
Adamantidis, Antoine ULg; Thomas, Elizabeth; Foidart, Agnès ULg et al

in European Journal of Neuroscience (2005), 21(10), 2837-44

In order to investigate the physiological properties of the melanin-concentrating hormone (MCH) we have generated and used mice from which the MCH receptor 1 gene was deleted (MCHR1(Neo/Neo) mice ... [more ▼]

In order to investigate the physiological properties of the melanin-concentrating hormone (MCH) we have generated and used mice from which the MCH receptor 1 gene was deleted (MCHR1(Neo/Neo) mice). Complementary experimental approaches were used to investigate alterations in the learning and memory processes of our transgenic model. The ability of the knockout strain to carry out the inhibitory passive avoidance test was found to be considerably impaired although no significant differences were observed in anxiety levels. This impaired cognitive property prompted us to explore modifications in N-methyl D-aspartate (NMDA) responses in the hippocampus. Intracellular recordings of CA1 pyramidal neurons in hippocampal slices from the MCHR1(Neo/Neo) mice revealed significantly decreased NMDA responses. Finally, using in situ hybridization we found a 15% reduction in NMDAR1 subunit in the CA1 region. These results show for the first time a possible role for MCH in the control of the function of the NMDA receptor. [less ▲]

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See detailPromoter characterization of the mouse melanin-concentrating hormone receptor 1
Lakaye, Bernard ULg; Adamantidis, Antoine ULg; Coumans, Bernard ULg et al

in Biochimica et Biophysica Acta-Gene Structure and Expression (2004), 1678(1), 1-6

The gene encoding the mouse melanin-concentrating hormone receptor 1 was isolated and its structural organization and flanking regions were characterized. The 3' flanking region is marked by the presence ... [more ▼]

The gene encoding the mouse melanin-concentrating hormone receptor 1 was isolated and its structural organization and flanking regions were characterized. The 3' flanking region is marked by the presence of two polyadenylation signals but used with different frequencies. RNase protection and 5' rapid amplification of cDNA ends (RACE) identified multiple transcription initiation sites between -150 and -203 bp upstream of the ATG initiation codon. Functional analysis of deletion mutants reveals a cell independent transcriptional activity localized between nucleotide -305 and -589. The proximal 1.5 kb region does not possess consensus TATA or CAAT boxes but has several consensus sequences for regulatory elements including USF, GATA, AP1, AP4, MyoD, GKLF and Ikaros that could explain the broad expression of the receptor. [less ▲]

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See detailNeuronal localization of the 25-kDa specific thiamine triphosphatase in rodent brain
Czerniecki, Jan ULg; Chanas, Grazyna; Verlaet, Myriam ULg et al

in Neuroscience (2004), 125(4), 833-840

Thiamine triphosphate (ThTP) is found in small amounts in most organisms from bacteria to mammals, but little is known about its physiological role. In vertebrate tissues, ThTP may act as a phosphate ... [more ▼]

Thiamine triphosphate (ThTP) is found in small amounts in most organisms from bacteria to mammals, but little is known about its physiological role. In vertebrate tissues, ThTP may act as a phosphate donor for the phosphorylation of certain proteins; this may be part of a new signal transduction pathway. We have recently characterized a highly specific 25-kDa thiamine triphosphatase (ThTPase) that is expressed in most mammalian tissues. The role of this enzyme may be the control of intracellular concentrations of ThTP. As the latter has been considered to be a neuroactive form of thiamine, we have studied the distribution of ThTPase mRNA and protein in rodent brain using in situ hybridization and immunohistochemistry. With both methods, we found the strongest staining in hippocampal pyramidal neurons, as well as cerebellar granule cells and Purkinje cells. Some interneurons were also labeled and many ThTPase mRNA-positive and immunoreactive cells were distributed throughout cerebral cortical gray matter and the thalamus. White matter was not significantly labeled. ThTPase immunoreactivity seems to be located mainly in the cytoplasm of neuronal perikarya. Immunocytochemical data using dissociated cultured cells from hippocampal and cerebellum showed that the staining was more intense in neurons than in astrocytes. The protein was rather uniformly located in the perikarya and dendrites, suggesting that ThTP and ThTPase may play a general role in neuronal metabolism rather than a specific role in excitability. There was no apparent correlation between ThTPase expression and selective vulnerability of certain brain regions to thiamine deficiency. (C) 2004 IBRO. Published by Elsevier Ltd. All rights reserved. [less ▲]

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See detailExpression of 25 kDa thiamine triphosphatase in rodent tissues using quantitative PCR and characterization of its mRNA
Lakaye, Bernard ULg; Verlaet, Myriam ULg; Dubail, Johanne ULg et al

in International Journal of Biochemistry & Cell Biology (2004), 36(10), 2032-2041

Thiamine triphosphate (ThTP) is found in most organisms, but its biological role remains unclear. In mammalian tissues, cellular ThTP concentrations remain low, probably because of hydrolysis by a ... [more ▼]

Thiamine triphosphate (ThTP) is found in most organisms, but its biological role remains unclear. In mammalian tissues, cellular ThTP concentrations remain low, probably because of hydrolysis by a specific 25 kDa thiamine triphosphatase (ThTPase). The aim of the present study was to use quantitative PCR, for comparing the 25 kDa ThTPase mRNA expression in various mouse tissues with its enzyme activities. ThTPase mRNA was expressed at only a few copies per cell. The highest amount of mRNA was found in testis, followed by lung and muscle, while the highest enzyme activities were found in liver and kidney. The poor correlation between mRNA levels and enzyme activities might result either from tissue-specific post-transcriptional regulation of mRNA processing and/or translation or from the regulation of enzyme activities by post-translational mechanisms. Purified recombinant human ThTPase was phosphorylated by casein kinase 11, but this phosphorylation did not modify the enzyme activity. However, the characterization of the 3'-untranslated mRNA region revealed a unique, highly conserved, 200-nucleotide sequence that might be involved in translational control. In situ hybridization studies in testis suggest a predominant localization of ThTPase mRNA in poorly differentiated spermatogenic cells. This is the first study demonstrating a cell-specific 25 kDa ThTPase mRNA expression, suggesting that this enzyme might be related to the degree of differentiation or the metabolic state of the cell. (C) 2004 Elsevier Ltd. All rights reserved. [less ▲]

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See detailHuman recombinant thiamine triphosphatase: purification, secondary structure and catalytic properties
Lakaye, Bernard ULg; Makarchikov, Alexander F; Wins, Pierre et al

in International Journal of Biochemistry & Cell Biology (2004), 36(7), 1348-1364

Thiamine triphosphate (ThTP) is found in most living organisms and it may act as a phosphate donor for protein phosphorylation. We have recently cloned the cDNA coding for a highly specific mammalian 25 ... [more ▼]

Thiamine triphosphate (ThTP) is found in most living organisms and it may act as a phosphate donor for protein phosphorylation. We have recently cloned the cDNA coding for a highly specific mammalian 25 kDa thiamine triphosphatase (ThTPase; EC 3.6.1.28). As the enzyme has a high catalytic efficiency and no sequence homology with known phosphohydrolases, it was worth investigating its structure and catalytic properties. For this purpose, we expressed the untagged recombinant human ThTPase (hThTPase) in E. coli, produced the protein on a large scale and purified it to homogeneity. Its kinetic properties were similar to those of the genuine human enzyme, indicating that the recombinant hThTPase is completely functional. Mg2+ ions were required for activity and Ca2+ inhibited the enzyme by competition with Mg2+. With ATP as substrate, the catalytic efficiency was 10(-4)-fold lower than with ThTP, confirming the nearly absolute specificity of the 25 kDa ThTPase for ThTP. The activity was maximum at pH 8.5 and very low at pH 6.0. Zn2+ ions were inhibitory at micromolar concentrations at pH 8.0 but activated at pH 6.0. Kinetic analysis suggests an activator site for Mg2+ and a separate regulatory site for Zn2+. The effects of group-specific reagents such as Woodward's reagent K and diethylpyrocarbonate suggest that at least one carboxyl group in the active site is essential for catalysis, while a positively charged amino group may be involved in substrate binding. The secondary structure of the enzyme, as determined by Fourier-transform infrared spectroscopy, was predominantly beta-sheet and alpha-helix. [less ▲]

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See detailThiamine triphosphate and thiamine triphosphatase activities: from bacteria to mammals
Makarchikov, Alexander F; Lakaye, Bernard ULg; Gulyai, I. E. et al

in Cellular and Molecular Life Sciences : CMLS (2003), 60(7), 1477-1488

In most organisms, the main form of thiamine is the coenzyme thiamine diphosphate. Thiamine triphosphate (ThTP) is also found in low amounts in most vertebrate tissues and can phosphorylate certain ... [more ▼]

In most organisms, the main form of thiamine is the coenzyme thiamine diphosphate. Thiamine triphosphate (ThTP) is also found in low amounts in most vertebrate tissues and can phosphorylate certain proteins. Here we show that ThTP exists not only in vertebrates but is present in bacteria, fungi, plants and invertebrates. Unexpectedly, we found that in Escherichia coli as well as in Arabidopsis thaliana, ThTP was synthesized only under particular circumstances such as hypoxia (E. coli) or withering (A. thaliana). In mammalian tissues, ThTP concentrations are regulated by a specific thiamine triphosphatase that we have recently characterized. This enzyme was found only in mammals. In other organisms, ThTP can be hydrolyzed by unspecific phosphohydrolases. The occurrence of ThTP from prokaryotes to mammals suggests that it may have a basic role in cell metabolism or cell signaling. A decreased content may contribute to the symptoms observed during thiamine deficiency. [less ▲]

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See detailBisindole alkaloids from Strychnos guianensis are effective antagonists of nicotinic acetylcholine receptors in cultured human TE671 cells
Wins, Pierre; Margineanu, Ilca; Penelle, Jacques et al

in Naunyn-Schmiedeberg's Archives of Pharmacology (2003), 367(3), 253-259

Several mono- and bisindole quaternary alkaloids isolated from the stem bark of Strychnos guianensis have recently been shown to be effective blockers of neuromuscular transmission in mice. In this study ... [more ▼]

Several mono- and bisindole quaternary alkaloids isolated from the stem bark of Strychnos guianensis have recently been shown to be effective blockers of neuromuscular transmission in mice. In this study, we used a human clonal cell line (TE671) expressing muscle-type nicotinic acetylcholine receptors. The agonist carbamylcholine activated a receptor-mediated Rb-86(+) efflux and this activation was antagonized by the indole alkaloids, the most active being bisindole bisquaternary compounds. The most effective antagonist, guiachrysine, had an IC50 around 0.43 muM in the presence of 0.5 mM carbamylcholine, compared to 0.16 muM for d-tubocurarine, the most potent curarizing alkaloid. Guiaflavine and 5',6'-dehydroguiaflavine were slightly less effective. Monoindole compounds were 10 to 100 times less potent than bisindole alkaloids. Kinetic analysis showed that the inhibition of the carbamylcholine-dependent Rb-86(+) efflux by guiaflavine was of mixed competitive and uncompetitive type. The competitive component (K-I = 0.21 muM) is presumably due to binding at the acetylcholine site, while the uncompetitive component (K'(I) = 0.92 muM) may be due to open channel block. [less ▲]

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See detailHuman immune cells express ppMCH mRNA and functional MCHR1 receptor
Verlaet, Myriam ULg; Adamantidis, Antoine ULg; Coumans, Bernard ULg et al

in FEBS Letters (2002), 527(1-3), 205-210

Melanin-concentrating hormone (MCH) is highly expressed in the brain and modulates feeding behavior. It is also expressed in some peripheral tissues where its role remains unknown. We have investigated ... [more ▼]

Melanin-concentrating hormone (MCH) is highly expressed in the brain and modulates feeding behavior. It is also expressed in some peripheral tissues where its role remains unknown. We have investigated MCH function in human and mouse immune cells. RT-PCR analysis revealed a low expression of prepro-MCH and MCH receptor 1 (MCHR1) but not of MCHR2 transcript in tissular and peripheral blood immune cells. FACS and in vitro assay studies demonstrated that MCHR1 receptor expression on most cell types can trigger, in the presence of MCH, cAMP synthesis and calcium mobilization in peripheral blood mononuclear cells (PBMCs). Moreover, MCH treatment decreases the CD3-stimulated PBMC proliferation in vitro. Accordingly, our data indicate for the first time that MCH and MCHR1 may exert immunomodulatory functions. (C) 2002 Federation of European Biochemical Societies. Published by Elsevier Science B.V. All rights reserved. [less ▲]

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See detailMolecular characterization of a specific thiamine triphosphatase widely expressed in mammalian tissues
Lakaye, Bernard ULg; Makarchikov, Alexander F; Antunes, Adelio F et al

in Journal of Biological Chemistry (2002), 277(16), 13771-13777

Thiamine triphosphate (ThTP) is found at low concentrations in most animal tissues, and recent data suggest that it may act as a phosphate donor for the phosphorylation of some proteins. In the mammalian ... [more ▼]

Thiamine triphosphate (ThTP) is found at low concentrations in most animal tissues, and recent data suggest that it may act as a phosphate donor for the phosphorylation of some proteins. In the mammalian brain, ThTP synthesis is rapid, but its steady-state concentration remains low, presumably because of rapid hydrolysis. In this report we purified a soluble thiamine triphosphatase (ThTPase; EC 3.6.1.28) from calf brain. The bovine ThTPase is a 24-kDa monomer, hydrolyzing ThTP with virtually absolute specificity. Partial sequence data obtained from the purified bovine enzyme by tandem mass spectrometry were used to search the GenBank(TM) data base. A significant identity was found with only one human sequence, the hypothetical 230-amino acid protein MGC2652. The coding regions from human and bovine brain mRNA were amplified by reverse transcription-PCR, cloned in Escherichia coli, and sequenced. The human open reading frame was expressed in E. coli as a GST fusion protein. Transformed bacteria had a high isopropyl-beta-D-thiogalactopyranoside-inducible ThTPase activity. The recombinant ThTPase had properties similar to those of human brain ThTPase, and it was specific for ThTP. The mRNA was expressed in most human tissues but at relatively low levels. This is the first report of a molecular characterization of a specific ThTPase. [less ▲]

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See detailAdenylate kinase 1 knockout mice have normal thiamine triphosphate levels
Makarchikov, Alexander F; Wins, Pierre; Janssen, E. et al

in Biochimica et Biophysica Acta-Molecular Cell Research (2002), 1592(2), 117-121

Thiamine triphosphate (ThTP) is found at low concentrations in most animal tissues and it may act as a phosphate donor for the phosphorylation of proteins, suggesting a potential role in cell signaling ... [more ▼]

Thiamine triphosphate (ThTP) is found at low concentrations in most animal tissues and it may act as a phosphate donor for the phosphorylation of proteins, suggesting a potential role in cell signaling. Two mechanisms have been proposed for the enzymatic synthesis of ThTP. A thiamine diphosphate (ThDP) kinase (ThDP + ATP double left right arrow ThTP + ADP) has been purified from brewer's yeast and shown to exist in rat liver. However, other data suggest that, at least in skeletal muscle, adenylate kinase I (AK1) is responsible for ThTP synthesis. In this study, we show that AK1 knockout mice have normal ThTP levels in skeletal muscle, heart, brain, liver and kidney, demonstrating that AK1 is not responsible for ThTP synthesis in those tissues. We predict that the high ThTP content of particular tissues like the Electrophorus electricus electric organ, or pig and chicken skeletal muscle is more tightly correlated with high ThDP kinase activity or low soluble ThTPase activity than with non-stringent substrate specificity and high activity of adenylate kinase. (C) 2002 Elsevier Science B.V All rights reserved. [less ▲]

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See detailATP-driven, Na(+)-independent inward Cl- pumping in neuroblastoma cells
Bettendorff, Lucien ULg; Lakaye, Bernard ULg; Margineanu, Ilca et al

in Journal of Neurochemistry (2002), 81(4), 792-801

In immature neurones, the steady-state intracellular Cl- concentration [Cl-](i) is generally higher than expected for passive distribution, and this is believed to be due to Na(+)-K(+)-2Cl(-) co-transport ... [more ▼]

In immature neurones, the steady-state intracellular Cl- concentration [Cl-](i) is generally higher than expected for passive distribution, and this is believed to be due to Na(+)-K(+)-2Cl(-) co-transport. Here, we show that N2a neuroblastoma cells, incubated in HEPES-buffered NaCl medium maintain a [Cl-](i) around 60 mm, two- to threefold higher than expected for passive distribution at a membrane potential of - 49 mV. When the cells were transferred to a Cl(-) -free medium, [Cl-](i) decreased quickly (t(1/2) < 5 min), suggesting a high Cl- permeability. When the intracellular ATP concentration was reduced to less than 1 mm by metabolic inhibitors, the initial rate of (36) Cl- uptake was strongly inhibited (60-65%) while steady-state [Cl-](i) decreased to 24 mm, close to the value predicted from the Nernst equilibrium. Moreover, after reduction of [ATP](i) and [Cl-](i) by rotenone, the subsequent addition of glucose led to a reaccumulation of Cl-, in parallel with ATP recovery. Internal bicarbonate did not affect Cl- pumping, suggesting that Cl-/HCO(3)(-) exchange does not significantly contribute to active transport. Likewise, Na(+) -K(+) -2Cl(-) co-transport also appeared to play a minor role: although mRNA for the NKCC1 form of the co-transporter was detected in N2a cells, neither the initial rate of (36)Cl- uptake nor steady-state [Cl-](i) were appreciably decreased by 10 microm bumetanide or replacement of external Na(+) by choline. These results suggest that a highly active ATP-dependent mechanism, distinct from Na(+) -K(+) -2Cl(-) co-transport, is responsible for most of the inward Cl- pumping in N2a cells. [less ▲]

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See detailThe genetic absence epilepsy rat from Strasbourg (GAERS), a rat model of absence epilepsy: Computer modeling and differential gene expression
Lakaye, Bernard ULg; Thomas, E.; Minet, Arlette ULg et al

in Epilepsia (2002), 43(Suppl. 5), 123-129

Purpose: We present results obtained by computer modeling of the thalamic network and differential gene expression analysis in a rat strain with absence epilepsy, the genetic absence epilepsy rat from ... [more ▼]

Purpose: We present results obtained by computer modeling of the thalamic network and differential gene expression analysis in a rat strain with absence epilepsy, the genetic absence epilepsy rat from Strasbourg (GAERS). Methods: (a) Computer modeling used equations from the Hodgking-Huxley model with a circuit of 13 reticular thalamic (nRt) and 39 thalamocortical (TC) neurons; (b) gene-expression analysis using differential mRNA display (DD), in situ hybridization, Northern blotting, and competitive reverse transcriptase-polymerase chain reaction (RT-PCR). Results: (a) Computer modeling showed an increased network synchrony in the thalamic circuit as the value of conductance of low-voltage activated calcium channel (LVACC) is increased. (b) Using differential mRNA display, a 40% upregulation of the H-ferritin mRNA in the hippocampus was demonstrated. Looking for some candidate genes of the VACC family, no difference was found in the alpha1G mRNA expression between GAERS and control animals, whereas a decreased expression of the alpha1E subunit was observed in the cerebellum and the brainstem of the GAERS. This phenomenon was not observed in young animals when the epileptic phenotype is not expressed. Conclusions: The use of computer modeling appeared to be an efficient way to evaluate the impacts of electrophysiologic findings in vivo from single cells on an entire circuit. No clear single gene defect was revealed so far in GAERS. More information could arise from linkage analysis. However, some brain structures like hippocampus or cerebellum classically not known to be involved in the production of absence spike-and-wave discharges could in fact participate in the development of this phenotype. [less ▲]

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See detailIncreased Expression of mRNA Encoding Ferritin Heavy Chain in Brain Structures of a Rat Model of Absence Epilepsy
Lakaye, Bernard ULg; de Borman, B.; Minet, Arlette ULg et al

in Experimental Neurology (2000), 162(1), 112-20

Differential mRNA display was carried out to find genes that are differentially regulated in the brain of a rat strain with absence epilepsy, the genetic absence epilepsy rats from Strasbourg (GAERS ... [more ▼]

Differential mRNA display was carried out to find genes that are differentially regulated in the brain of a rat strain with absence epilepsy, the genetic absence epilepsy rats from Strasbourg (GAERS). Among the 32 differentially displayed cDNA fragments actually cloned and sequenced, one shows 100% identity with the rat heavy chain ferritin (H-ferritin) mRNA. Northern blot analysis confirmed the up-regulation of the H-ferritin mRNA. Using dot blotting, a 40% increase in expression was reported in the subcortical forebrain of the adult GAERS, while cortex, brain stem, and cerebellum appeared unmodified. This change was not observed in the brain of 25-day-old rats, an age at which the epileptic phenotype is not present. By in situ hybridization, the enhanced expression was localized in the hippocampus. The increase in mRNA encoding H-ferritin was not immunodetected at the protein level by Western blotting. These results are not apparently related to the neural substrate of SWD or to the distribution of local increase in glucose metabolism previously described in the GAERS. It is hypothesized that the up-regulation of the H-ferritin mRNA is part of a mechanism protecting the hippocampus, a seizure-prone area, against a possible overactivation during absence seizures. [less ▲]

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See detailEffect of nicotine on rat gingival fibroblasts in vitro
Lahmouzi, Jamila ULg; Simain-Sato, Franklin ULg; Defresne, Marie-Paule ULg et al

in Connective Tissue Research (2000), 41(1), 69-80

Nicotine from 3 to 5 mM affects growth and survival rate of rat gingival fibroblasts in vitro. Ultrastructural analysis revealed dilated mitochondria and vacuolization in treated cells, suggestive of ... [more ▼]

Nicotine from 3 to 5 mM affects growth and survival rate of rat gingival fibroblasts in vitro. Ultrastructural analysis revealed dilated mitochondria and vacuolization in treated cells, suggestive of necrosis, but increased apoptosis was also revealed by cytometry. On the basis of this in vitro study, it appear that tobacco, through its component nicotine, may directly affect various functions of rat gingival fibroblasts [less ▲]

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See detailCulture of Gingival Fibroblasts on Bioabsorbable Regenerative Materials in Vitro
Simain-Sato, Franklin ULg; Lahmouzi, Jamila ULg; Kalykakis, G. K. et al

in Journal of Periodontology (1999), 70(10), 1234-9

BACKGROUND: The use of membranes in guided tissue regeneration (GTR) can limit the apical migration of gingival cells and favor the establishment of new attachment by periodontal ligament fibroblasts ... [more ▼]

BACKGROUND: The use of membranes in guided tissue regeneration (GTR) can limit the apical migration of gingival cells and favor the establishment of new attachment by periodontal ligament fibroblasts. However, gingival recession during healing following GTR has been described as a frequent complication. The purpose of this study was to determine if gingival fibroblasts are affected by the composition of the bioabsorbable membranes used in mucogingival surgery. METHODS: Two type of bioabsorbable regenerative materials were used as cell carriers. Wistar rat gingival fibroblasts (RGF) were obtained from attached gingiva, cut into small fragments, and placed in culture dishes. When confluent, cells were detached using trypsin and identified as "first transferred cells" (P1). At the third passage (P3), cell count, trypan blue exclusion test, acid phosphatase activity, DNA synthesis, phase contrast microscopy, and scanning electron microscopy were performed. The cells were then placed in wells containing the membranes and incubated for 72 hours. RESULTS: When examined under microscopy, the control wells (without membranes) showed one cell type with the elongated appearance characteristic of fibroblasts. The wells with membranes showed an altered cell morphology with a high proportion of cell fragments regardless of the type of membrane used. CONCLUSIONS: These results suggest that cell carrier membranes could affect RGF morphology and thus alter gingival tissue healing following GTR. [less ▲]

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See detailEstrogen Receptor-Beta in Quail: Cloning, Tissue Expression and Neuroanatomical Distribution
Foidart, Agnès ULg; Lakaye, Bernard ULg; Grisar, Thierry ULg et al

in Journal of Neurobiology (1999), 40(3), 327-42

A partial estrogen receptor-beta (ERbeta) cDNA had been previously cloned and sequenced in Japanese quail. The 3'- and 5'-rapid amplification of cDNA ends techniques were used here to identify a cDNA ... [more ▼]

A partial estrogen receptor-beta (ERbeta) cDNA had been previously cloned and sequenced in Japanese quail. The 3'- and 5'-rapid amplification of cDNA ends techniques were used here to identify a cDNA sequence of the quail ERbeta that contains a complete open reading frame. For the first time in an avian species, this cDNA sequence and the corresponding amino acid sequence are described. They are compared with the known ERbeta sequences previously described in mammals and with the ERalpha sequences identified in a selection of mammalian and avian species. The analysis by Northern blotting of the ERbeta mRNA expression in the brain and kidneys revealed the presence of several transcripts. The presence of ERbeta identified by reverse transcriptase-polymerase chain reaction demonstrated a widespread distribution quite different from the distribution of ERalpha. The complete neuroanatomical distribution of ERbeta mRNA as determined by in situ hybridization with 35S- and 33P-labeled oligoprobes is also presented. Transcripts are present in many nuclei implicated in the control of reproduction such as the medial preoptic nucleus, the nucleus striae terminalis, and the nucleus taeniae, the avian homologue of the amygdala. These data demonstrate the presence of ERbeta in a nonmammalian species and indicate that the (neuro)-anatomical distribution of this receptor type has been conserved in these two classes of vertebrates. The role of this receptor in the control of reproduction and other physiological processes should now be investigated. [less ▲]

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See detailExpression of Mrna Encoding Alpha1e and Alpha1g Subunit in the Brain of a Rat Model of Absence Epilepsy
de Borman, B.; Lakaye, Bernard ULg; Minet, Arlette ULg et al

in Neuroreport (1999), 10(3), 569-74

Low voltage-activated calcium channels are thought to play a key role in the generation of spike and waves discharges characteristic of absence epilepsy. Therefore, the expression level of mRNA encoding ... [more ▼]

Low voltage-activated calcium channels are thought to play a key role in the generation of spike and waves discharges characteristic of absence epilepsy. Therefore, the expression level of mRNA encoding calcium channel alpha1E and alpha1G subunits was measured in the brain of genetic absence epilepsy rats from Strasbourg (GAERS). Using quantitative RT-PCR and in situ hybridization, no difference was found in alpha1G mRNA expression between GAERS and control animals, while a decreased expression of alpha1E was seen in the cerebellum and the brain stem of the GAERS. This phenomenon was not observed in young animals when the epileptic phenotype is not expressed. [less ▲]

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See detailThe Molecular Neuron-Glia Couple and Epileptogenesis
Grisar, Thierry ULg; Lakaye, Bernard ULg; Thomas, E. et al

in Advances in Neurology (1999), 79

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