GPR88, coding for a G protein-coupled orphan receptor that is highly represented in the striatum,... more GPR88, coding for a G protein-coupled orphan receptor that is highly represented in the striatum, is a strong functional candidate gene for neuropsychiatric disorders and is located at 1p22-p21, a chromosomal region that we have previously linked to bipolar disorder (BD) in the Sardinian population. In order to ascertain the relevance of GPR88 as a risk factor for psychiatric diseases, we performed a genetic association analysis between GPR88 and BD in a sample of triads (patient and both parents) recruited in the Sardinian and the Palestinian population as well as between GPR88 and schizophrenia (SZ) in triads from the Xhosa population in South Africa. We found a positive association between GPR88 and BD in the Sardinian and Palestinian triads. Moreover, we found a positive association between GPR88 and SZ in triads from the Xhosa population in South Africa. When these results were corrected for multiple testing, the association between GPR88 and BD was maintained in the Palestinian population. Thus, these results suggest that GPR88 deserves consideration as a candidate gene for psychiatric diseases and requires to be further investigated in other populations.
Activity-dependent regulation of Egr1/Zif268, a transcription factor (TF) of the Egr family, is e... more Activity-dependent regulation of Egr1/Zif268, a transcription factor (TF) of the Egr family, is essential for stabilization of dentate gyrus synaptic plasticity and consolidation and reconsolidation of several forms of memory. The gene can be rapidly induced in selective brain circuits after certain types of learning or after recall. Here, we focused on area CA1 and examined regulation of Egr1, Egr2, and Egr3 mRNA and protein, and their DNA binding activity to the Egr response element (ERE) at different times after LTP in vivo and after learning and recall of a fear memory. We found LTP in CA1 leads to rapid induction of the three Egrs, however only Egr1 protein was overexpressed without a co-ordinated change in binding activity, indicating a fundamental difference between CA1 and dentate gyrus LTP. Our investigations in fear memory reveal that both learning and retrieval lead to an increase in binding of constitutively expressed Egr1 and Egr3 to the ERE, but not Egr2. Memory recall was also associated with increased Egr1 protein translation. The nature and temporal dynamics of these changes and tests for interactions between TFs suggest that in addition to ERE-mediated transcription, Egr1 in CA1 may interact with the TF c-Fos to regulate genes via other DNA response elements. V V C 2011 Wiley Periodicals, Inc.
Transformation of a steroidogenic mouse adrenal cell line (Y-I) by simian adenovirus SA7 produced... more Transformation of a steroidogenic mouse adrenal cell line (Y-I) by simian adenovirus SA7 produced a ceil line with low apparent steroidogenic activity. The effect of ACTH and cholera toxin on cyclic AMP production was similar in both not transformed and virus-transformed cells and activity of cyclic AMP=dependent protein kinase wa% also similar in both ceils. In transformed ceils, cholesterol was metabolized to A'-3fS-hydroxysteroids, mainly 20c~-~ihydropregnenolone while in not transformed ceils, the major metabolites were A -3 ketosteroids (20~-dihydroand ll[~-hydroxy-20a-dihydroprogesterone). In both cell lines ACTH increased the metabolism of cholesterol. Further studies with labelled pregnenolone and progesterone revealed a loss of A -3[3-hydroxysteroid dehydrogenase/isomerase and I I[~-hydroxylase activity in the transformed cells.
Two strains of cytomegalovirus (CMV) from patients with renal transplants and two other strains o... more Two strains of cytomegalovirus (CMV) from patients with renal transplants and two other strains of CMV from young children were successfully propagated in human diploid cells, MRC5. Analysis by SDS-PAGE of these viruses passaged only 4 to 6 times revealed 21 to 29 polypeptides. The freshly isolated strains differed from reference strains, Ad 169 and Davis, in the number of bands, in the intensity of labeling of each band and also in molecular weight of the polypeptides. Some bands were common to all six strains. Some bands of molecular weight above 105 K were present in some strains and not in others. This was also the case for some bands between 64 and 76 Kand two to five bands at low molecular weight between 17 and 22 K were detected. The band at 68 K was the most intensely labeled. Several polypeptides were precipitated by immune sera containing antibodies to CMV (one positive human serum and one guinea pig antiserum to Davis strain). Different bands were observed after precipitation, depending on the antiserum used and the strain of virus. Interstrain differences were found to be associated with antigenic heterogeneity. However, these cannot be correlated as yet with primary infection or reactivation.
Increased synaptic stimulation of sympathetic neurons in vivo causes a delayed increase in the ac... more Increased synaptic stimulation of sympathetic neurons in vivo causes a delayed increase in the activity and the amount of tyrosine hydroxylase (TH). To determine whether these changes result from an increase in the messenger RNA for TH, the rat preganglionic cervical sympathetic trunk was electrically stimulated unilaterally for 90 min, and 48 h later RNA was extracted from stimulated and contralateral control superior cervical ganglia. Northern blots probed with a cDNA for TH demonstrated that nerve stimulation produced about a 2.5-fold increase in the amount of TH mRNA in the ganglion. These results indicate that synaptic stimulation leads to an increase in TH mRNA, either by increasing the rate of transcription of the TH gene or by increasing the stability of its mRNA.
Idiopathic Parkinson&... more Idiopathic Parkinson's disease (PD) is characterized by mesencephalic dopaminergic neuron cell death and striatal dopamine (DA) depletion. The factors involved in the pathogenesis of the disease are still unknown. Transforming growth factor beta1 (TGFbeta1) is increased in the striatum of patients with PD. However, the effect of this increase is not known. Here, we show that overexpression of TGFbeta1, by recombinant adenovirus TGFbeta1 gene transfer, in the mesostriatal system of an MPTP mouse model of PD decreased the number of mesencephalic dopaminergic neurons. This effect also involved more extensive DA depletion in the striatum. Striatal enkephalin mRNA levels are augmented, suggesting a decrease in dopaminergic transmission to the postsynaptic target. In the absence of MPTP, TGFbeta1 greatly decreased the number of dopaminergic neurons in the ventral mesencephalon of fully mature mice. These results show that an increase in TGFbeta1 levels aggravate the parkinsonian status of MPTP mice and may therefore be a risk factor for the development of PD.
... Rolando Meloni, Jacques Mallet, and Nicole Faucon Biguet ... For instance, micro-implants act... more ... Rolando Meloni, Jacques Mallet, and Nicole Faucon Biguet ... For instance, micro-implants acting as sensors and signal transductors have proven to be a valid therapeutic tool for hearing loss (Bishop and Eby, 2010; Peterson et al, 2010) and are now trialled to re-establish vision ...
Microsatellites are common repeated sequences, which are useful as genetic markers and lack any c... more Microsatellites are common repeated sequences, which are useful as genetic markers and lack any clearly established function. In a previous study we suggested that an intronic polymorphic TCAT repeat in the tyrosine hydroxylase (TH) gene, the microsatellite HUMTH01, may regulate transcription. The TH gene encodes the rate-limiting enzyme in the synthesis of catecholamines, and the microsatellite HUMTH01 has been used in genetic studies of neuropsychiatric and cardiovascular diseases, in which disturbances of catecholaminergic neurotransmission have been implicated. HUMTH01 alleles associated with these diseases act as transcriptional enhancers when linked to a minimal promoter and are recognized by specific nuclear factors. Here we show that allelic variations of HUMTH01 commonly found in humans have a quantitative silencing effect on TH gene expression. Two specific proteins, ZNF191, a zinc finger protein, and HBP1, an HMG box transcription factor, which bind the TCAT motif, were then cloned. Finally, allelic variations of HUMTH01 correlate with quantitative and qualitative changes in the binding by ZNF191. Thus, this repeated sequence may contribute to the control of expression of quantitative genetic traits. As the HUMTH01 core motif is ubiquitous in the genome, this phenomenon may be relevant to the quantitative expression of many genes in addition to TH.
Background: Sarcopenia is a major public health problem in industrialized nations, placing an inc... more Background: Sarcopenia is a major public health problem in industrialized nations, placing an increasing burden on public healthcare systems because the loss of skeletal muscle mass and strength that characterizes this affection increases the dependence and the risk of injury caused by sudden falls in elderly people. Albeit exercise and caloric restriction improve sarcopenia-associated decline of the muscular performances, a more suitable and focused pharmacological treatment is still lacking.
Many studies provide evidence that retinoic acid (RA), an endogenous derivative of vitamin A, pla... more Many studies provide evidence that retinoic acid (RA), an endogenous derivative of vitamin A, plays a role in the development of the nervous system. We now report that RA controls the neurotransmitter phenotype of post-mitotic rat sympathetic neurons in cell culture. RA added to the culture medium increased the specific activity of choline acetyltransferase (ChAT) and the level of acetylcholine (ACh). Concomitantly, RA reduced the specific activities of two catecholamine synthetic enzymes, tyrosine hydroxylase (TH) and dopamine beta-hydroxylase (DBH) and the level of norepinephrine (NE). After a 2 week treatment with 5 microM RA, ChAT was increased by 5-10 fold, whereas TH and DBH were decreased by 10-15 fold and 2-3 fold, respectively, as compared to sympathetic neurons grown in the absence of RA. The modulation of the activity of the three enzymes was dose-dependent and followed a similar time course. The decrease of TH expression was demonstrated to be due to a decreased number of TH molecules.
The role played by protein kinase C (PKC) in TH gene regulation was investigated at transcription... more The role played by protein kinase C (PKC) in TH gene regulation was investigated at transcriptional and post-transcriptional levels using PC12 cells. The cells were treated with the phorbol ester TPA, which not only activates PKC but also causes down-regulation. PKC levels were monitored by [3H]PDBU binding assay and by using an anti-PKC antibody that detected intact PKC (79 kd) as well as its catalytic and regulatory domains. The [3H]PDBU binding to the membrane-associated PKC increased within 15-30 min of TPA treatment; thereafter total cellular [3H]PDBU binding decreased to a minimum of 20% of the control at 8 h. The rate of decrease in binding was greater than the decrease in the intensity of the staining of PKC holo enzyme visualized by anti-PKC antibody. TH mRNA levels, measured over the same time period, rose within 15 min of TPA treatment to peak at 4 h and subsequently declined below control level, paralleling the depletion of PKC. If cells depleted of PKC were reincubated ...
A single injection of reserpine causes a long lasting enhancement of the activity of tyrosine hyd... more A single injection of reserpine causes a long lasting enhancement of the activity of tyrosine hydroxylase (TH), the enzyme catalyzing the rate-limiting step in the biosynthesis of catecholamines. A sensitive method has been developed to assay both TH mRNA level and enzyme activity in tissue from a single rat. The time course of the induction was analysed in adrenals, locus coeruleus and substantia nigra. In both locus coeruleus and adrenals reserpine caused respectively 4.2- and 4.5-fold increase of TH mRNA which was maximal 2 days after drug injection. This increase is about twice that of the enzyme activity. No change was observed in substantia nigra. The effect lasted longer in locus coeruleus than in adrenal. In the latter, TH mRNA had almost returned to initial values at day 4 whereas at this time it is 3-fold higher in locus coeruleus and still significant at day 18. This result suggests that induction of TH results from an enhanced transcription of the TH gene. The time cours...
We have analyzed some functional aspects of the promoter of the human dopamine beta-hydroxylase (... more We have analyzed some functional aspects of the promoter of the human dopamine beta-hydroxylase (DBH) gene. A fragment of 1,247 bp directly 5' to the transcriptional start was progressively shortened, placed in front of a reporter gene, and tested in a human neuroblastoma cell line expressing DBH (SK-N-SH-TFM) and in a monkey kidney cell line (CV-1). A remarkably short region (267 bp), directly upstream from the transcription start, was sufficient to confer activity and tissue-specific expression. Furthermore, the expression of the DBH gene was shown to be inducible by cyclic AMP in SK-N-SH-TFM cells. This effect was demonstrated to occur at the transcriptional level, as shown by run-on assays, and was due to the presence of a near-consensus cyclic AMP-responsive element located in the untranscribed 5' regulatory region of the gene.
Comptes rendus de l'Académie des sciences. Série III, Sciences de la vie, 1984
The expression of tyrosine-hydroxylase (TH) gene was analysed in tissue sections of bovine adrena... more The expression of tyrosine-hydroxylase (TH) gene was analysed in tissue sections of bovine adrenal glands, by in situ hybridization using a single-stranded cDNA probe. Tissue fixation and hybridization conditions were found that led to a specific and sensitive detection of TH.
A full length dopamine-beta-hydroxylase (DBH) cDNA clone was isolated from a human pheochromocyto... more A full length dopamine-beta-hydroxylase (DBH) cDNA clone was isolated from a human pheochromocytoma lambda gt11 library. Both structural and functional evidence confirms the authenticity of the clone: (i) antibodies selected with fusion proteins generated by positive clones precipitate DBH activity, (ii) the sequence of three internal DBH tryptic peptides are included in the deduced DBH sequence, (iii) the previously reported N-terminal 15 amino acids of bovine DBH exhibits a nearly complete identity with that predicted for human DBH. The polypeptide chain of DBH comprises 578 amino acids corresponding to an unmodified protein of 64 862 daltons and is preceded by a cleaved signal peptide of 25 residues. DBH exists in both membrane-bound and soluble forms. The hydropathy plot reveals no obvious hydrophobic segment, except the signal peptide. S1 mapping analysis indicates no diversity in the 5' and 3' extremities of the DBH mRNA. Taken together with available biochemical data,...
Preganglionic nerve stimulation has been shown to induce delayed or long term changes in the neur... more Preganglionic nerve stimulation has been shown to induce delayed or long term changes in the neuron. Different models were used to study the trans-synaptic regulation of the tyrosine hydroxylase (TH): the rat adrenal medulla (AM) and the superior cervical ganglia (SCG). Northern blot analysis, western blot and enzymatic assays demonstrated that the TH mRNA paralleled both an increase in the protein amount and its enzymatic activity. Results of in vitro transcription on nuclei isolated from AM or from SCG after treatment with reserpine suggest that this increase in TH expression is due to an effect on the transcriptional activity of the TH gene. Other gene, cfos, is also induced by reserpine indicating that the TH transcription in these neurons may be mediated by "third messengers". Several putative regulatory elements, in particular an octamer sequence AP1 has been localized in the promoter of TH gene. Gel shift assays with nuclear extracts from untreated and phorbol ester...
The anterior pituitary is thought to be unable to synthesize dopamine (DA) except under experimen... more The anterior pituitary is thought to be unable to synthesize dopamine (DA) except under experimental conditions where a tyrosine hydroxylase (TH) activity, the rate-limiting step of its synthesis, has been demonstrated.
GPR88, coding for a G protein-coupled orphan receptor that is highly represented in the striatum,... more GPR88, coding for a G protein-coupled orphan receptor that is highly represented in the striatum, is a strong functional candidate gene for neuropsychiatric disorders and is located at 1p22-p21, a chromosomal region that we have previously linked to bipolar disorder (BD) in the Sardinian population. In order to ascertain the relevance of GPR88 as a risk factor for psychiatric diseases, we performed a genetic association analysis between GPR88 and BD in a sample of triads (patient and both parents) recruited in the Sardinian and the Palestinian population as well as between GPR88 and schizophrenia (SZ) in triads from the Xhosa population in South Africa. We found a positive association between GPR88 and BD in the Sardinian and Palestinian triads. Moreover, we found a positive association between GPR88 and SZ in triads from the Xhosa population in South Africa. When these results were corrected for multiple testing, the association between GPR88 and BD was maintained in the Palestinian population. Thus, these results suggest that GPR88 deserves consideration as a candidate gene for psychiatric diseases and requires to be further investigated in other populations.
Activity-dependent regulation of Egr1/Zif268, a transcription factor (TF) of the Egr family, is e... more Activity-dependent regulation of Egr1/Zif268, a transcription factor (TF) of the Egr family, is essential for stabilization of dentate gyrus synaptic plasticity and consolidation and reconsolidation of several forms of memory. The gene can be rapidly induced in selective brain circuits after certain types of learning or after recall. Here, we focused on area CA1 and examined regulation of Egr1, Egr2, and Egr3 mRNA and protein, and their DNA binding activity to the Egr response element (ERE) at different times after LTP in vivo and after learning and recall of a fear memory. We found LTP in CA1 leads to rapid induction of the three Egrs, however only Egr1 protein was overexpressed without a co-ordinated change in binding activity, indicating a fundamental difference between CA1 and dentate gyrus LTP. Our investigations in fear memory reveal that both learning and retrieval lead to an increase in binding of constitutively expressed Egr1 and Egr3 to the ERE, but not Egr2. Memory recall was also associated with increased Egr1 protein translation. The nature and temporal dynamics of these changes and tests for interactions between TFs suggest that in addition to ERE-mediated transcription, Egr1 in CA1 may interact with the TF c-Fos to regulate genes via other DNA response elements. V V C 2011 Wiley Periodicals, Inc.
Transformation of a steroidogenic mouse adrenal cell line (Y-I) by simian adenovirus SA7 produced... more Transformation of a steroidogenic mouse adrenal cell line (Y-I) by simian adenovirus SA7 produced a ceil line with low apparent steroidogenic activity. The effect of ACTH and cholera toxin on cyclic AMP production was similar in both not transformed and virus-transformed cells and activity of cyclic AMP=dependent protein kinase wa% also similar in both ceils. In transformed ceils, cholesterol was metabolized to A'-3fS-hydroxysteroids, mainly 20c~-~ihydropregnenolone while in not transformed ceils, the major metabolites were A -3 ketosteroids (20~-dihydroand ll[~-hydroxy-20a-dihydroprogesterone). In both cell lines ACTH increased the metabolism of cholesterol. Further studies with labelled pregnenolone and progesterone revealed a loss of A -3[3-hydroxysteroid dehydrogenase/isomerase and I I[~-hydroxylase activity in the transformed cells.
Two strains of cytomegalovirus (CMV) from patients with renal transplants and two other strains o... more Two strains of cytomegalovirus (CMV) from patients with renal transplants and two other strains of CMV from young children were successfully propagated in human diploid cells, MRC5. Analysis by SDS-PAGE of these viruses passaged only 4 to 6 times revealed 21 to 29 polypeptides. The freshly isolated strains differed from reference strains, Ad 169 and Davis, in the number of bands, in the intensity of labeling of each band and also in molecular weight of the polypeptides. Some bands were common to all six strains. Some bands of molecular weight above 105 K were present in some strains and not in others. This was also the case for some bands between 64 and 76 Kand two to five bands at low molecular weight between 17 and 22 K were detected. The band at 68 K was the most intensely labeled. Several polypeptides were precipitated by immune sera containing antibodies to CMV (one positive human serum and one guinea pig antiserum to Davis strain). Different bands were observed after precipitation, depending on the antiserum used and the strain of virus. Interstrain differences were found to be associated with antigenic heterogeneity. However, these cannot be correlated as yet with primary infection or reactivation.
Increased synaptic stimulation of sympathetic neurons in vivo causes a delayed increase in the ac... more Increased synaptic stimulation of sympathetic neurons in vivo causes a delayed increase in the activity and the amount of tyrosine hydroxylase (TH). To determine whether these changes result from an increase in the messenger RNA for TH, the rat preganglionic cervical sympathetic trunk was electrically stimulated unilaterally for 90 min, and 48 h later RNA was extracted from stimulated and contralateral control superior cervical ganglia. Northern blots probed with a cDNA for TH demonstrated that nerve stimulation produced about a 2.5-fold increase in the amount of TH mRNA in the ganglion. These results indicate that synaptic stimulation leads to an increase in TH mRNA, either by increasing the rate of transcription of the TH gene or by increasing the stability of its mRNA.
Idiopathic Parkinson&... more Idiopathic Parkinson's disease (PD) is characterized by mesencephalic dopaminergic neuron cell death and striatal dopamine (DA) depletion. The factors involved in the pathogenesis of the disease are still unknown. Transforming growth factor beta1 (TGFbeta1) is increased in the striatum of patients with PD. However, the effect of this increase is not known. Here, we show that overexpression of TGFbeta1, by recombinant adenovirus TGFbeta1 gene transfer, in the mesostriatal system of an MPTP mouse model of PD decreased the number of mesencephalic dopaminergic neurons. This effect also involved more extensive DA depletion in the striatum. Striatal enkephalin mRNA levels are augmented, suggesting a decrease in dopaminergic transmission to the postsynaptic target. In the absence of MPTP, TGFbeta1 greatly decreased the number of dopaminergic neurons in the ventral mesencephalon of fully mature mice. These results show that an increase in TGFbeta1 levels aggravate the parkinsonian status of MPTP mice and may therefore be a risk factor for the development of PD.
... Rolando Meloni, Jacques Mallet, and Nicole Faucon Biguet ... For instance, micro-implants act... more ... Rolando Meloni, Jacques Mallet, and Nicole Faucon Biguet ... For instance, micro-implants acting as sensors and signal transductors have proven to be a valid therapeutic tool for hearing loss (Bishop and Eby, 2010; Peterson et al, 2010) and are now trialled to re-establish vision ...
Microsatellites are common repeated sequences, which are useful as genetic markers and lack any c... more Microsatellites are common repeated sequences, which are useful as genetic markers and lack any clearly established function. In a previous study we suggested that an intronic polymorphic TCAT repeat in the tyrosine hydroxylase (TH) gene, the microsatellite HUMTH01, may regulate transcription. The TH gene encodes the rate-limiting enzyme in the synthesis of catecholamines, and the microsatellite HUMTH01 has been used in genetic studies of neuropsychiatric and cardiovascular diseases, in which disturbances of catecholaminergic neurotransmission have been implicated. HUMTH01 alleles associated with these diseases act as transcriptional enhancers when linked to a minimal promoter and are recognized by specific nuclear factors. Here we show that allelic variations of HUMTH01 commonly found in humans have a quantitative silencing effect on TH gene expression. Two specific proteins, ZNF191, a zinc finger protein, and HBP1, an HMG box transcription factor, which bind the TCAT motif, were then cloned. Finally, allelic variations of HUMTH01 correlate with quantitative and qualitative changes in the binding by ZNF191. Thus, this repeated sequence may contribute to the control of expression of quantitative genetic traits. As the HUMTH01 core motif is ubiquitous in the genome, this phenomenon may be relevant to the quantitative expression of many genes in addition to TH.
Background: Sarcopenia is a major public health problem in industrialized nations, placing an inc... more Background: Sarcopenia is a major public health problem in industrialized nations, placing an increasing burden on public healthcare systems because the loss of skeletal muscle mass and strength that characterizes this affection increases the dependence and the risk of injury caused by sudden falls in elderly people. Albeit exercise and caloric restriction improve sarcopenia-associated decline of the muscular performances, a more suitable and focused pharmacological treatment is still lacking.
Many studies provide evidence that retinoic acid (RA), an endogenous derivative of vitamin A, pla... more Many studies provide evidence that retinoic acid (RA), an endogenous derivative of vitamin A, plays a role in the development of the nervous system. We now report that RA controls the neurotransmitter phenotype of post-mitotic rat sympathetic neurons in cell culture. RA added to the culture medium increased the specific activity of choline acetyltransferase (ChAT) and the level of acetylcholine (ACh). Concomitantly, RA reduced the specific activities of two catecholamine synthetic enzymes, tyrosine hydroxylase (TH) and dopamine beta-hydroxylase (DBH) and the level of norepinephrine (NE). After a 2 week treatment with 5 microM RA, ChAT was increased by 5-10 fold, whereas TH and DBH were decreased by 10-15 fold and 2-3 fold, respectively, as compared to sympathetic neurons grown in the absence of RA. The modulation of the activity of the three enzymes was dose-dependent and followed a similar time course. The decrease of TH expression was demonstrated to be due to a decreased number of TH molecules.
The role played by protein kinase C (PKC) in TH gene regulation was investigated at transcription... more The role played by protein kinase C (PKC) in TH gene regulation was investigated at transcriptional and post-transcriptional levels using PC12 cells. The cells were treated with the phorbol ester TPA, which not only activates PKC but also causes down-regulation. PKC levels were monitored by [3H]PDBU binding assay and by using an anti-PKC antibody that detected intact PKC (79 kd) as well as its catalytic and regulatory domains. The [3H]PDBU binding to the membrane-associated PKC increased within 15-30 min of TPA treatment; thereafter total cellular [3H]PDBU binding decreased to a minimum of 20% of the control at 8 h. The rate of decrease in binding was greater than the decrease in the intensity of the staining of PKC holo enzyme visualized by anti-PKC antibody. TH mRNA levels, measured over the same time period, rose within 15 min of TPA treatment to peak at 4 h and subsequently declined below control level, paralleling the depletion of PKC. If cells depleted of PKC were reincubated ...
A single injection of reserpine causes a long lasting enhancement of the activity of tyrosine hyd... more A single injection of reserpine causes a long lasting enhancement of the activity of tyrosine hydroxylase (TH), the enzyme catalyzing the rate-limiting step in the biosynthesis of catecholamines. A sensitive method has been developed to assay both TH mRNA level and enzyme activity in tissue from a single rat. The time course of the induction was analysed in adrenals, locus coeruleus and substantia nigra. In both locus coeruleus and adrenals reserpine caused respectively 4.2- and 4.5-fold increase of TH mRNA which was maximal 2 days after drug injection. This increase is about twice that of the enzyme activity. No change was observed in substantia nigra. The effect lasted longer in locus coeruleus than in adrenal. In the latter, TH mRNA had almost returned to initial values at day 4 whereas at this time it is 3-fold higher in locus coeruleus and still significant at day 18. This result suggests that induction of TH results from an enhanced transcription of the TH gene. The time cours...
We have analyzed some functional aspects of the promoter of the human dopamine beta-hydroxylase (... more We have analyzed some functional aspects of the promoter of the human dopamine beta-hydroxylase (DBH) gene. A fragment of 1,247 bp directly 5' to the transcriptional start was progressively shortened, placed in front of a reporter gene, and tested in a human neuroblastoma cell line expressing DBH (SK-N-SH-TFM) and in a monkey kidney cell line (CV-1). A remarkably short region (267 bp), directly upstream from the transcription start, was sufficient to confer activity and tissue-specific expression. Furthermore, the expression of the DBH gene was shown to be inducible by cyclic AMP in SK-N-SH-TFM cells. This effect was demonstrated to occur at the transcriptional level, as shown by run-on assays, and was due to the presence of a near-consensus cyclic AMP-responsive element located in the untranscribed 5' regulatory region of the gene.
Comptes rendus de l'Académie des sciences. Série III, Sciences de la vie, 1984
The expression of tyrosine-hydroxylase (TH) gene was analysed in tissue sections of bovine adrena... more The expression of tyrosine-hydroxylase (TH) gene was analysed in tissue sections of bovine adrenal glands, by in situ hybridization using a single-stranded cDNA probe. Tissue fixation and hybridization conditions were found that led to a specific and sensitive detection of TH.
A full length dopamine-beta-hydroxylase (DBH) cDNA clone was isolated from a human pheochromocyto... more A full length dopamine-beta-hydroxylase (DBH) cDNA clone was isolated from a human pheochromocytoma lambda gt11 library. Both structural and functional evidence confirms the authenticity of the clone: (i) antibodies selected with fusion proteins generated by positive clones precipitate DBH activity, (ii) the sequence of three internal DBH tryptic peptides are included in the deduced DBH sequence, (iii) the previously reported N-terminal 15 amino acids of bovine DBH exhibits a nearly complete identity with that predicted for human DBH. The polypeptide chain of DBH comprises 578 amino acids corresponding to an unmodified protein of 64 862 daltons and is preceded by a cleaved signal peptide of 25 residues. DBH exists in both membrane-bound and soluble forms. The hydropathy plot reveals no obvious hydrophobic segment, except the signal peptide. S1 mapping analysis indicates no diversity in the 5' and 3' extremities of the DBH mRNA. Taken together with available biochemical data,...
Preganglionic nerve stimulation has been shown to induce delayed or long term changes in the neur... more Preganglionic nerve stimulation has been shown to induce delayed or long term changes in the neuron. Different models were used to study the trans-synaptic regulation of the tyrosine hydroxylase (TH): the rat adrenal medulla (AM) and the superior cervical ganglia (SCG). Northern blot analysis, western blot and enzymatic assays demonstrated that the TH mRNA paralleled both an increase in the protein amount and its enzymatic activity. Results of in vitro transcription on nuclei isolated from AM or from SCG after treatment with reserpine suggest that this increase in TH expression is due to an effect on the transcriptional activity of the TH gene. Other gene, cfos, is also induced by reserpine indicating that the TH transcription in these neurons may be mediated by "third messengers". Several putative regulatory elements, in particular an octamer sequence AP1 has been localized in the promoter of TH gene. Gel shift assays with nuclear extracts from untreated and phorbol ester...
The anterior pituitary is thought to be unable to synthesize dopamine (DA) except under experimen... more The anterior pituitary is thought to be unable to synthesize dopamine (DA) except under experimental conditions where a tyrosine hydroxylase (TH) activity, the rate-limiting step of its synthesis, has been demonstrated.
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Papers by Nicole Biguet