Valérie Metzinger-Le Meuth | Université Sorbonne Paris Nord / Sorbonne Paris Nord University (original) (raw)

Papers by Valérie Metzinger-Le Meuth

1 INRA-Agrocampus, UMR de génétique animale, 35042 Rennes Cedex. 2 Présente adresse Université Pa... more 1 INRA-Agrocampus, UMR de génétique animale, 35042 Rennes Cedex. 2 Présente adresse Université Paris 13, 93017 Bobigny Cedex. AGENAE-chick, un outil d'analyse du transcriptome de la poule Dans le cadre du programme d'analyse du génome des animaux d'élevage (AGENAE) une banque d'ADNc multi-tissus normalisée (collection de fragments d'ADN correspondant à des gènes exprimés) a été construite pour l'espèce poule. Plus de 13000 clones ont ainsi été obtenus. Leur séquençage a permis d'obtenir 23273 séquences partielles (EST pour étiquette de séquence exprimée) publiées dans la base de données publiques de l'EMBL. Ces séquences correspondent à environ 6800 gènes exprimés différents. Leur analyse effectuée grâce à des outils développés par l'équipe informatique SIGENAE a permis de confirmer la représentativité des différents tissus utilisés pour la construction de la banque. Elle a aussi montré la bonne représentativité des fonctions moléculaires associée...

AJP Gastrointestinal and Liver Physiology

HPLC-purified 125I-labeled vasoactive intestinal peptide (VIP) bound in a specific, saturable, an... more HPLC-purified 125I-labeled vasoactive intestinal peptide (VIP) bound in a specific, saturable, and reversible manner to pancreatic plasma membranes isolated from newborn calves, from milk-fed calves at 28 and 119 days, and from weaned calves at 119 days. A series of VIP analogues, including pituitary adenylate cyclase-activating polypeptide (PACAP), displaced 125I-VIP binding and activated adenylate cyclase in the same order of relative potency: PACAP-38 greater than helodermin greater than VIP, PACAP-27 greater than PHM (human peptide with NH2-terminal histidine and COOH-terminal methionine amide). At maximally effective concentrations, these five peptides produced the same two- to threefold increase of adenylate cyclase activity in pancreatic membranes from newborn and 28-day-old calves, and fourfold in ruminant or preruminant animals at 119 days. The activation constant for PACAP-38 ranged from 0.1 to 0.34 nM throughout the postnatal development. Helospectin I and II were three times less potent than VIP in inhibiting 125I-VIP binding. At concentrations up to 0.1 microM, secretin, rat and human growth hormone-releasing factors, glucagon, oxyntomodulin, the truncated form of glucagon-like peptide-1 lacking the 6 NH2-terminal amino acid sequence (TGLP-1), GLP-2, gastric inhibitory peptide, gastrin, CCK, and insulin had no effect on binding. Scatchard plots from 28- and 119-day-old calves were compatible with the presence of two classes of 125I-VIP binding sites: one with a high affinity for VIP and a low binding capacity (Kd = 0.11-0.4 nM, Bmax = 66-174 fmol/mg protein) and the other with a low affinity and high binding capacity. At birth, only one class of binding sites was observed (Kd = 0.4 nM, Bmax = 858 fmol/mg protein). The covalently cross-linked PACAP-preferring 125I-VIP binding site is a glycoprotein of 55 kDa with higher sensitivity to PACAP vs. helodermin and VIP. Our results suggest that calf pancreatic functions might be regulated at an early stage of postnatal development by PACAP receptors linked to cAMP generation.

Résumé L'INRA a initié un programme de recherche génomique (AGENAE) pour 4 espèces animales, ... more Résumé L'INRA a initié un programme de recherche génomique (AGENAE) pour 4 espèces animales, dont la poule. L'un des axes de ce programme concerne la génomique fonctionnelle, et, dans ce cadre, une banque multitissus d'ADNc (collection d'ADNc correspondant aux gènes exprimés) de l'espèce poule a été réalisée. Plus de 14000 clones ont été séquencés et sont disponibles pour des études d'expression de gènes. Les premières utilisations de ces outils, dans le cadre de recherches sur le contrôle du sexe et la maîtrise de la composition corporelle dans l'espèce poule sont présentés.

e347. doi: 10.14800/rd.347; © 2014 by Metzinger-Le Meuth V, et al. Development of disease is ofte... more e347. doi: 10.14800/rd.347; © 2014 by Metzinger-Le Meuth V, et al. Development of disease is often due to deregulation of gene expression. The gene program is controlled at the post-transcriptional level by the action of small non-coding RNAs known as microRNAs (miRNAs), short, single-stranded molecules that control mRNA stability or translational repression via base pairing with regions in the 3' untranslated region of their target mRNAs. Over the last decade, considerable progress has been made to elucidate the roles of miRNAs in vascular pathogenesis and develop the use of miRNAs as innovative biomarkers in diagnostics, and as groundbreaking drugs in pharmacological treatments. It has been recently shown that several miRNAs are implicated in the course of chronic kidney disease (CKD) and are associated with vessel damage, such as vascular calcifications and atherosclerosis. The inflammatory miR-223 is increased in vitro in vascular smooth muscle cells subjected to uremic toxi...

BMC molecular biology, Jan 7, 2002

Specific cis-elements and the associated trans-acting factors have been implicated in the post-tr... more Specific cis-elements and the associated trans-acting factors have been implicated in the post-transcriptional regulation of gene expression. In the era of genome wide analyses identifying novel trans-acting factors and cis-regulatory elements is a step towards understanding coordinated gene expression. UV-crosslink analysis is a standard method used to identify RNA-binding proteins. Uridine is traditionally used to radiolabel substrate RNAs, however, proteins binding to cis-elements particularly uridine poor will be weakly or not detected. We evaluate here the possibility of using UV-crosslinking with RNA substrates radiolabeled with each of the four ribonucleotides as an approach for screening for novel sequence specific RNA-binding proteins. The radiolabeled RNA substrates were derived from the 3'UTRs of the cloned Eg and c-mos Xenopus laevis maternal mRNAs. Specific, but not identical, uv-crosslinking signals were obtained, some of which corresponded to already identified pr...

Cancer metastasis reviews, 1997

The recent advances in the understanding of the pathogenesis of ovarian cancer have been helpful ... more The recent advances in the understanding of the pathogenesis of ovarian cancer have been helpful in addressing issues in diagnosis, prognosis and management. The study of ovarian tumours by novel techniques such as immunohistochemistry, fluorescent in situ hybridisation, comparative genomic hybridisation, polymerase chain reaction and new tumour markers have aided the evaluation and application of new concepts into clinical practice. The correlation of novel surrogate tumour specific features with response to treatment and outcome in patients has defined prognostic factors which may allow the future design of tailored therapy based on a molecular profile of the tumour. These have also been used to design new approaches to therapy such as antibody targeting and gene therapy. The delineation of roles of c-erbB2, c-fms and other novel receptor kinases in the pathogenesis of ovarian cancer has led initially to the development of anti-c-erbB2 monoclonal antibody therapy. The discovery of...

Frontiers in Bioscience, 2014

Vascular calcification arises during chronic kidney disease (CKD), and increases the risk of card... more Vascular calcification arises during chronic kidney disease (CKD), and increases the risk of cardiovascular mortality. In CKD, alterations of cerebral circulation were linked with an increase in ischemic strokes and behavioral troubles. Studying pathophysiological mechanisms of calcifications and detecting new biomarkers in the cerebral circulation is thus an important issue. microRNAs are small non-coding, single-stranded RNAs that regulate messenger RNAs at the post-transcriptional level. They are involved in numerous pathologies and represent new opportunities to develop disease predictors. We used RT-qPCR to quantify endothelial-specific microRNAs in cerebral arterioles from WT mice and from pathological models of CKD. We used four mice groups: WT SHAM, WT CKD, Apolipoprotein E Knock-Out (ApoE-KO) SHAM, ApoE-KO CKD. Brains were removed after two and ten weeks of uremia and RNA from cerebral arterioles was extracted. miR-17 and miR-126 were the most dysregulated in the pathological conditions, at both the second week and tenth week of uremia. Our results suggest that miR-17 and miR-126 are potential new biomarkers of cerebral troubles of CKD patients and new therapeutic targets for innovative treatments.

Biochimica et Biophysica Acta (BBA) - Molecular Basis of Disease, 2014

a b s t r a c t 8 MicroRNAs (miRNAs) are small, noncoding RNAs of 18-22 nucleotides in length tha... more a b s t r a c t 8 MicroRNAs (miRNAs) are small, noncoding RNAs of 18-22 nucleotides in length that regulate post-22 transcriptional expression by base-pairing with target mRNAs. It is now clearly established that miRNAs are 23 involved in most of the cell's physiopathological processes (including carcinogenesis and metabolic disorders).

Current Basic and Pathological Approaches to the Function of Muscle Cells and Tissues - From Molecules to Humans, 2012

ABSTRACT In the last 10 years, microRNAs (miRNAs) have emerged as critical regulators of numerous... more ABSTRACT In the last 10 years, microRNAs (miRNAs) have emerged as critical regulators of numerous physiological and pathological mechanisms [1-2], including cardiac and vascular smooth muscle cell (VSMC) plasticity [3-5]. These small molecules (approx. 20 to 25 nucleotides) comprise a novel and abundant class of endogenous interfering RNAs. More than 1 500 miRNAs are now listed by dedicated internet databases such as miRBase, Tarbase, MicroRNA.org or miRdb (See sub-chapter II for URL adresses). They are transcribed and matured, in a process known as miRNA biogenesis [6] which starts with the transcription of a larger RNA product, called pri-miRNA, by the RNA polymerase II in the vast majority of cases. Pri-miRNA, which is a few hundred to a few thousand nucleotides long, is then submitted to cleavage in the nucleus by a specific RNase III (Drosha) and its protein partner, DiGeorge syndrome critical region 8 (DGCR8), near the base of the miRNA hairpin stem. This process releases a pre-miRNA hairpin (of approx. 60 to 70 nt). The pre-miRNA is then released in the cytoplasm where it is recognized and cleaved within its stems by the Dicer RNase III and its protein partners. This results in a double stranded RNA, known as the miRNA/miRNA* duplex (approx. 22 bp). This complex is unwound to single strands. One strand (guiding strand / mature miRNA) is incorporated in the RNA-induced-silencing complex (RISC) that contains Argonaute 2 (Ago2), another endonuclease, the other strand is usually rapidly degraded. Finally, the RISC complex carries the mature miRNA to its target messenger RNAs (mRNAs), which results in gene silencing, in a post-transcriptional manner [7]. Figure 1 shows a representative example of the biogenesis of miR-143 and miR-145, which are the main miRNAs expressed in smooth muscle cells.

PLoS ONE, 2012

Ashr a f Y usu f Ra n gre z 1,2 . ¤ , Elé o n or e M'B a y a-M o u t o ula 1,2 . , V alé rie M e ... more Ashr a f Y usu f Ra n gre z 1,2 . ¤ , Elé o n or e M'B a y a-M o u t o ula 1,2 . , V alé rie M e t zin g er-Le M e u t h 1,4" ,

Pancreas, 2005

Secretin is a key regulator of pancreatic secretion, but the molecular basis of its action is not... more Secretin is a key regulator of pancreatic secretion, but the molecular basis of its action is not well understood, especially in the calf pancreas. Our study investigated the expression and functional competence of secretin receptors (SEC-R) in calf pancreatic membranes. We used reverse transcriptase-polymerase chain reaction, sequencing, and Northern blot to assess the expression of the SEC-R gene. The functional characterization of SEC-R was accomplished using adenylate cyclase (AC) assay. We successfully amplified, by reverse transcriptase-polymerase chain reaction, a fragment of the SEC-R gene from 119-day-old calf pancreas. This sequence shows higher homology with SEC-R than with vasoactive intestinal polypeptide (VIP)-1 and VIP-2 receptors from other species. Northern blot analysis detected a 1.8-kb transcript. Accordingly, secretin stimulates AC activity in calf pancreatic membranes isolated from 28- and 119-day-old animals with a potency (Ka) of 1.9 to 2.7 nmol/L. Maximal AC stimulation induced by secretin represented a 3- to 4-fold increase of basal activity. AC activation by secretin was inhibited by the 2 SEC-R antagonists, [psi4,5] secretin (l micromol/L) and [5-27] secretin (10 micromol/L). Interestingly, [psi4,5] secretin was ineffective against VIP-induced AC stimulation. Our data indicate that secretin exerts a direct action on pancreatic secretion through specific SEC-R coupled to the AC system. Calf pancreatic SEC-Rs are coexpressed with VIP-2 receptors that we previously identified by ligand binding and cross-linking experiments.

Oncogene, 2002

We have previously shown a high frequency of allele loss at D6S193 (62%) on chromosomal arm 6q27 ... more We have previously shown a high frequency of allele loss at D6S193 (62%) on chromosomal arm 6q27 in ovarian tumours and mapped the minimal region of allele loss between D6S297 and D6S264 (3 cM). We isolated and mapped a single non-chimaeric YAC (17IA12, 260 ± 280 kb) containing D6S193 and D6S297. A further extended bacterial contig (between D6S264 and D6S149) has been established using PACs and BACs and a transcript map has been established. We have mapped six new markers to the YAC; three of them are ESTs (WI-15078, WI-8751, and TCP10). We have isolated three cDNA clones of EST WI-15078 and one clone contains a complete open reading frame. The sequence shows homology to a new member of the ribonuclease family. The other two clones are splice variants of this new gene. The gene is expressed ubiquitously in normal tissues. It is expressed in 4/8 ovarian cancer cell lines by Northern analysis. The gene encodes for a 40 kDa protein. Direct sequencing of the gene in all the eight ovarian cancer cell lines did not identify any mutations. Clonogenic assays were performed by transfecting the full-length gene in to ovarian cancer cell lines and no suppression of growth was observed.

Nutrition Research Reviews, 2006

The aim of the present review is to synthesise and summarise our recent knowledge on the involvem... more The aim of the present review is to synthesise and summarise our recent knowledge on the involvement of cholecystokinin (CCK) and gastrin peptides and their receptors in the control of digestive functions and more generally their role in the field of nutrition in mammals. First, we examined the release of these peptides from the gut, focusing on their molecular forms, the factors regulating their release and the signalling pathways mediating their effects. Second, general physiological effects of CCK and gastrin peptides are described with regard to their specific receptors and the role of CCK on vagal mucosal afferent nerve activities. Local effects of CCK and gastrin in the gut are also reported, including gut development, gastrointestinal motility and control of pancreatic functions through vagal afferent pathways, including NO. Third, some examples of the intervention of the CCK and gastrin peptides are exposed in diseases, taking into account intervention of the classical receptor subtypes (CCK 1 and CCK 2 receptors) and their heterodimerisation as well as CCK-C receptor subtype. Finally, applications and future challenges are suggested in the nutritional field (performances) and in therapy with regards to the molecular forms or in relation with the type of receptor as well as new techniques to be utilised in detection or in therapy of disease. In conclusion, the present review underlines recent developments in this field: CCK and gastrin peptides and their receptors are the key factor of nutritional aspects; a better understanding of the mechanisms involved may increase the efficiency of the nutritional functions and the treatment of abnormalities under pathological conditions. Nutrition: Gastrin: Cholecystokinin: Cholecystokinin -gastrin family receptors: Digestive physiology: Digestive diseases

JBIC Journal of Biological Inorganic Chemistry, 2013

In addition to its electron transfer activity, cytochrome c is now known to trigger apoptosis via... more In addition to its electron transfer activity, cytochrome c is now known to trigger apoptosis via peroxidase activity. This new function is related to a structural modification of the cytochrome upon association with anionic lipids, particularly cardiolipin present in the mitochondrial membrane. However, the exact nature of the non-native state induced by this interaction remains an active subject of debate. In this work, using human cytochromes c (native and two single-histidine mutants and the corresponding double mutant) and micelles as a hydrophobic medium, we succeeded, through UV-visible spectroscopy, circular dichroism spectroscopy and NMR spectroscopy, in fully characterizing the nature of the sixth ligand replacing the native methionine. Furthermore, careful pH titrations permitted the identification of the amino acids involved in the iron binding over a range of pH values. Replacement of the methionine by lysine was only observed at pH above 8.5, whereas histidine binding is dependent on both pH and micelle concentration. The pH variation range for histidine protonation is relatively narrow and is consistent with the mitochondrial intermembrane pH changes occurring during apoptosis. These results allow us to rule out lysine as the sixth ligand at pH values close to neutrality and reinforce the role of histidines (preferentially His33 vs. His26) as the main candidate to replace methionine in the non-native cytochrome c. Finally, on the basis of these results and molecular dynamics simulations, we propose a 3D model for non-native cytochrome c in a micellar environment.

Circulation: Cardiovascular Genetics, 2011

Biochimica et Biophysica Acta (BBA) - Molecular Basis of Disease, 2014

disease (CKD) is associated with vascular calcifications and atherosclerosis. There is a need for... more disease (CKD) is associated with vascular calcifications and atherosclerosis. There is a need for 28 novel predictors to allow earlier diagnosis of these disorders, predict disease progression, and improve 29 assessment of treatment response. We focused on microRNAs since they are implicated in a variety of cellular 30 functions in cardiovascular pathology. We examined changes of microRNA expression in aortas of CKD and 31 non-CKD wild type mice and apolipoprotein E knock-out mice, respectively. Both vascular smooth muscle-32 specific miR-143 and miR-145 expressions were decreased in states of atherosclerosis and/or CKD or both, and 33 the expression level of protein target Myocardin was increased. The inflammatory miR-223 was increased in 34 more advanced stages of CKD, and specific protein targets NFI-A and GLUT-4 were dramatically decreased. 35 Expression of miR-126 was markedly increased and expression of protein targets VCAM-1 and SDF-1 was altered 36 during the course of CKD. The drug sevelamer, commonly used in CKD, corrected partially these changes in 37 microRNA expression, suggesting a direct link between the observed microRNA alterations and uremic vascular 38 toxicity. Finally, miR-126, -143 and -223 expression levels were deregulated in murine serum during the course 39 of experimental CKD. In conclusion, these miRNAs could have role(s) in CKD vascular remodeling and may 40 therefore represent useful targets to prevent or treat complications of CKD. 41 42 43 44 45 46 65 course of experimental CKD and CKD-associated atherosclerosis has 66 not been studied so far. We therefore decided to analyze the expression 67 of six cardiovascular-related miRNAs [9] in aortas from murine models 68 with CKD, atherosclerosis, and vascular calcification [10]. Wild type 69 (WT) C57BL/6J as well as Apolipoprotein-E knock-out (Apo-E KO) 70 mice underwent partial nephrectomy to induce CKD. Apo-E KO mice 71 characteristically develop large atheromatous plaques and low-grade 72 vascular calcification. The combination of ApoE-KO and CKD leads to 73 more marked atherosclerosis and aortic calcification than in Apo-E KO Biochimica et Biophysica Acta xxx (2013) xxx-xxx

AJP Gastrointestinal and Liver Physiology

BMC molecular biology, Jan 7, 2002

Cancer metastasis reviews, 1997

Frontiers in Bioscience, 2014

Biochimica et Biophysica Acta (BBA) - Molecular Basis of Disease, 2014

Current Basic and Pathological Approaches to the Function of Muscle Cells and Tissues - From Molecules to Humans, 2012

PLoS ONE, 2012

Pancreas, 2005

Oncogene, 2002

Nutrition Research Reviews, 2006

JBIC Journal of Biological Inorganic Chemistry, 2013

Circulation: Cardiovascular Genetics, 2011

Biochimica et Biophysica Acta (BBA) - Molecular Basis of Disease, 2014