Biomedical subjects

Y Vandenbrouck

Publications and source records attributed to Y Vandenbrouck.

5 recordsLinked to original sources

PubMed1999

The Genexpress IMAGE knowledge base of the human brain transcriptome: a prototype integrated resource for functional and computational genomics.

Expression profiles of 5058 human gene transcripts represented by an array of 7451 clones from the first IMAGE Consortium cDNA library from infant brain have been collected by semiquantitative hybridization of the array with complex probes derived by reverse transcription of mRNA from brain and five other human tissues. Twenty-one percent of the clones corresponded to transcripts that could be classified in general categories of low, moderate, or high abundance. These expression profiles were integrated with cDNA clone and sequence clustering and gene mapping information from an upgraded version of the Genexpress Index. For seven gene transcripts found to be transcribed preferentially or specifically in brain, the expression profiles were confirmed by Northern blot analyses of mRNA from eight adult and four fetal tissues, and 15 distinct regions of brain. In four instances, further documentation of the sites of expression was obtained by in situ hybridization of rat-brain tissue sections. A systematic effort was undertaken to further integrate available cytogenetic, genetic, physical, and genic map informations through radiation-hybrid mapping to provide a unique validated map location for each of these genes in relation to the disease map. The resulting Genexpress IMAGE Knowledge Base is illustrated by five examples presented in the printed article with additional data available on a dedicated Web site at the address http://idefix.upr420.vjf.cnrs.fr/EXPR++ +/ welcome.html.

Brain Chemistry↗

PubMed1996

Transcriptional regulation of apolipoprotein E expression by cyclic AMP.

Incubation of HepG2 cells in the presence of dibutyryl cAMP (db-cAMP), a cell permeable analogue of cyclic AMP, or forskolin, an agent which elevates intracellular cAMP, resulted in a 50% decrease in apoE mRNA levels within 24 h. Results of nuclear run-on transcription assays showed that db-cAMP down-regulates apoE gene expression at the transcriptional level. By transfection analysis with a plasmid containing the -614/+804 human apoE gene fused to the secreted placental alkaline phosphatase (SPAP) reporter gene, we showed that the SPAP activity was decreased by 50% when HepG2 cells were incubated in the presence of db-cAMP or forskolin, indicating that this promoter region mediated this negative effect. In contrast, when the smaller fragment -200/+1 of apoE promoter was linked to the CAT reporter gene, db-cAMP treatment of HepG2 cells resulted in a 2-fold increase in CAT activity, suggesting that positive cAMP-responsive elements were present in the proximal apoE promoter. These data indicate that transcriptional modulation of apoE gene expression by agents known to elevate the intracellular cAMP level is complex and involves several negative and positive elements located in the -614 to +804 region of the apoE gene whose global effect is negative on apoE gene transcription.

Apolipoproteins E↗

PubMed1995

Transcriptional regulation of apolipoprotein A-I expression in Hep G2 cells by phorbol ester.

The regulation of apolipoprotein A-I (apo A-I) gene expression by 12-O-tetradecanoylphorbol 13-acetate (TPA) was investigated in the human hepatoma cell line Hep G2. TPA treatment decreased apo A-I mRNA levels in a time-dependent manner, by up to 50% versus control cells within 24 h. Nuclear run-on transcription assays demonstrated a transcriptional effect of TPA. Using transfection analysis with a plasmid construct containing the -1378/+11 apo A-I promoter fused to the secreted placental alkaline phosphatase (SPAP) reporter gene, we showed that the SPAP activity was decreased to 50% when Hep G2 cells were incubated in the presence of TPA. The inhibitory effect of TPA was still maintained when fragment -253 to -4 of apo A-I promoter was linked to the CAT reporter gene. These data indicate that transcriptional modulation of apolipoprotein A-I gene expression following phorbol ester treatment is transduced by gene elements located between -253 and -4 of the apo A-I promoter.

Apolipoprotein A-I↗

PubMed1995

Thyroid hormone modulates apolipoprotein-AI gene expression at the post-transcriptional level in Hep G2 cells.

Hyperthyroidism is associated with elevated plasma levels of apolipoprotein AI (apo AI). We have examined the effects of 3,3',-5-triiodothyronine on apo AI mRNA, transcription run-on activity, apo AI mRNA half-life, and the rate of protein synthesis in Hep G2 cells, to understand the molecular mechanism by which thyroid hormone regulates apo AI gene expression. Incubation with thyroid hormone increased the apo AI and apo AII mRNA concentrations twofold. Cycloheximide alone caused a significant increase in apo AI mRNA. Nuclear run-on assays indicate that thyroid hormone did not change the rate of the apo AI gene transcription at 6, 12 or 24 h, showing that thyroid hormone did not modulate apo AI gene transcription. Kinetic studies performed in the presence of actinomycin D showed that the half-life of apo AI mRNA was increased 2-3-fold by thyroid hormone over control cells. Thyroid hormone did not change the incorporation of [35S]methionine into immunoprecipitable apo AI. Pulse-chase experiments demonstrated that there was no change in the secretion and degradation rates of labeled apo AI in response to T3. This suggests that thyroid hormone does not affect the catabolism of apo AI (degradation or/and uptake) and that translation control strongly influences the regulation of apo AI gene expression. The stabilization of apo AI mRNA by thyroid hormone and its role in translation remain to be elucidated.

Apolipoprotein A-I↗

PubMed1994

The modulation of apolipoprotein E gene expression by 3,3'-5-triiodothyronine in HepG2 cells occurs at transcriptional and post-transcriptional levels.

The regulation of the synthesis and secretion of apolipoprotein E (apoE) is incompletely understood. This study examines the mechanisms responsible for regulating apoE gene expression in HepG2 cells by thyroid hormone (3,3'-5-triiodothyronine). The secretion rate of apoE was by thyroid hormone increased (1.5-1.8-fold) in pulse/chase experiments. Thyroid hormone doubled apoE mRNA concentration as determined by Northern-blot analysis. Inhibition of protein synthesis by cycloheximide increased the thyroid-hormone-induced stimulation of apoE mRNA. This suggests that the synthesis of new protein is not required for thyroid hormone to stimulate apoE mRNA. Actinomycin D was used to inhibit new transcription; there was a more rapid degradation of mature apoE mRNA in thyroid hormone-treated HepG2 cells than in control cells, suggesting that thyroid hormone acts post-transcriptionally to regulate apoE gene expression. Cycloheximide blocked the action of thyroid hormone, suggesting that thyroid hormone regulates the turnover of apoE mRNA via the synthesis of de novo protein. Nuclear run-on transcription assays demonstrated that thyroid hormone stimulated apoE gene transcription threefold in 24 h. These findings indicate that the expression of the apoE gene is controlled at both transcriptional and post-transcriptional loci by the thyroid hormone.

Apolipoproteins E↗