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P Clerc

Publications and source records attributed to P Clerc.

At least 19 recordsLinked to original sources
PubMed2001

Tsix-mediated repression of Xist accumulation is not sufficient for normal random X inactivation.

During the X inactivation process, one X chromosome in each female embryonic cell is chosen at random to become coated by Xist RNA and silenced. Tsix, a transcript anti-sense to Xist, participates in the choice of the inactive X and in Xist regulation through as yet unknown mechanisms. Undifferentiated female ES cells, which have two active Xs, recapitulate random X inactivation when induced to differentiate. A 65 kb deletion targeted to one of the two Xs in a female ES cell line, and including both the end of the Xist gene and the site of initiation of Tsix, resulted in the exclusive inactivation of the deleted X in differentiated ES cells. We have re-examined the phenotype of the 65 kb deletion and targeted Tsix and the terminal exons of Xist back to the deleted locus using a cre/loxP site-specific re-insertion strategy. We show that prior to inactivation the deleted X is associated in undifferentiated ES cells with both increased Xist expression and diffusion of the Xist transcript away from its site of synthesis. Restoration of Tsix repressed the steady-state level of Xist expression and restricted Xist RNA to its transcription site. At the onset of inactivation in differentiated ES cells, restoration of Tsix failed to restore random X-inactivation, even though the levels of Xist RNA accumulation in cis were markedly reduced. These results identify for the first time a dual function for Tsix as both a repressor of the steady-state level of Xist expression and as a regulator of the distribution of Xist RNA within the nucleus. They also establish that random inactivation requires mechanisms additional to the in cis repression of XIST:

Alleles↗
PubMed2000

Genetics. Reprogramming X inactivation.

Inactivation of one of the two X chromosomes occurs in all cells of female adult mice so that genes are expressed from only one X chromosome. In a Perspective, Clerc and Avner describe an elegant series of experiments in mouse embryos cloned from adult and embryonic female cell nuclei (Eggan et al.) that reveal how the inactivation state of the X chromosomes is reprogrammed.

Animals↗
PubMed2000

Usefulness of three-dimensional echocardiography for the evaluation of mitral valve prolapse: an intraoperative study.

BACKGROUND AND AIM OF THE STUDY: The study aim was to evaluate the feasibility of intraoperative three-dimensional (3D) transesophageal echocardiography (TEE) in patients referred for mitral valve prolapse (MVP) repair and to compare two-dimensional (2D) TEE and 3D TEE and surgical findings. METHODS: Forty-six patients (mean age 67 +/- 11 years) underwent 3D TEE intraoperatively. Measurements were made of the posterior part of mitral annulus circumference (PMAC), and the width of mitral valve surgical resection on the mitral annulus (WMVR). Using 3D TEE, MVP topography was described, and PMAC in diastole and the width of implantation of MVP on the mitral annulus (WMVP) in systole were measured. RESULTS: 3D TEE was successful in 42 patients (91%). 2D and 3DTEE correctly predicted MVP localization in 38 (90%) and 36 (86%) patients, respectively (p = NS). 3D TEE and surgical PMAC were 89 +/- 13 and 93 +/- 21 mm, respectively (p = 0.01, R = 0.42). WMVR and WMVP were 28 +/- 11 mm and 26 +/- 11 mm, respectively (p <0.0001, R = 0.82). WMVR/anatomic PMAC (0.29 +/- 0.11) and WMVP/3D echo PMAC (0.32 +/- 0.11) were correlated (p <0.0001, R= 0.69). CONCLUSION: Intraoperative 3D TEE evaluation of MVP is feasible. MVP width and its ratio to the mitral annulus were assessed, and found to correlate with surgical findings. These 3D data may be of value to the surgeon when performing mitral valve repair.

Aged↗
PubMed1999

The CCKB/gastrin receptor is coupled to the regulation of enzyme secretion, protein synthesis and p70 S6 kinase activity in acinar cells from ElasCCKB transgenic mice.

In order to determine which physiological functions can be regulated by the pancreatic CCKB/gastrin receptor, studies were carried out on pancreatic acini from mice expressing transgenic CCKB/gastrin receptors in the exocrine pancreas (ElasCCKB mice). Acini were stimulated by sulfated gastrin in the presence of SR 27897 (1.8 microM), blocking endogenous CCKA receptors. After 30 min incubation with gastrin, the secretion of chymotrypsinogen and amylase showed superimposable monophasic dose-response curves. Enzyme secretion was detectable and maximal at 100 pM and 1 nM of gastrin, respectively. No increase in chymotrypsinogen and amylase mRNAs was detected for doses of gastrin which specifically occupy the CCKB/gastrin receptor. In contrast, gastrin stimulated total protein synthesis in isolated acini from ElasCCKB mice. [35S]Methionine incorporation into total proteins was increased dose-dependently to a maximum for 30 pM gastrin and inhibited with higher doses (> 300 pM). Gastrin stimulated p70 S6 kinase activity for concentrations ranging from 10 pM to 1 nM. Gastrin-stimulated p70 S6 kinase activity and protein synthesis were blocked by rapamycin and wortmannin. Therefore, in ElasCCKB mice acinar cells, the CCKB/gastrin receptor mediates enzyme release and protein synthesis. However, a more efficient coupling of the CCKB/gastrin receptor to protein synthesis than to enzyme secretion was demonstrated. CCKB/gastrin receptor-stimulated protein synthesis likely results from an enhancement of mRNA translation and involves phosphatidyl inositol 3-kinase and p70 S6 kinase.

Amino Acid Sequence↗
PubMed1999

Evidence for a functional role of the cholecystokinin-B/gastrin receptor in the human fetal and adult pancreas.

Gastrin (G) and cholecystokinin (CCK) are gastrointestinal neuropeptides that are released into circulation during a meal. G is also transiently expressed during embryogenic and early ontogenic development of the pancreas and is believed to act on islet-cell development. Both peptides act on pancreatic endocrine function; however, the effects are dependent on the species and on cellular and molecular underlying mechanisms that remain poorly characterized. Since CCK-B/G subtype receptor is predominant over the CCK-A subtype in the human pancreas, we hypothesized that it could be expressed by islet cells. Here we present reverse transcription-polymerase chain reaction and immunohistochemistry data demonstrating that the CCK-B/G receptor is expressed in islet cells and that islet glucagon-producing cells are the major site of CCK-B/G receptor expression in adult and fetal pancreas. Moreover, G immunoreactivity was detected in the fetal human pancreas at embryogenic week 22. G- and CCK-stimulated glucagon are released from purified human islets. Concentration of CCK and G eliciting a half-maximal level of glucagon secretion were 13 +/- 6 and 8 +/- 5 pmol/l, respectively. Maximal glucagon secretion was achieved in the presence of 30 pmol/l peptides and was similar to that obtained in the presence of 10 mmol/l L-arginine (1.6 pmol x ml(-1) x 90 min(-1)). The nonpeptide antagonist of the CCK-B/G receptor, RPR-101048, fully inhibited CCK- and G-stimulated glucagon secretion at 100 nmol/l concentration. These data are consistent with the view that the CCK-B/G receptor is involved in glucose homeostasis in adult humans and mediates the autocrine effects of G on islet differentiation and growth in the fetal pancreas.

Adult↗
PubMed1998

Transgenic CCK-B/gastrin receptor mediates murine exocrine pancreatic secretion.

BACKGROUND & AIMS: The presence of cholecystokinin (CCK)-B/gastrin receptors in the pancreas of higher mammals including humans has been shown, but their physiological function in the normal pancreas is unknown. The aim of this study was to investigate whether they couple to the secretory machinery of normal acinar cells. METHODS: A transgenic mouse strain expressing the human CCK-B/gastrin receptor in the exocrine pancreas was created. The transgenic construction used the promoter region of the elastase I gene and the human CCK-B/gastrin receptor gene. Analysis of ElasCCKB mice included polymerase chain reaction and receptor autoradiography. Molecular and binding features of the CCK-B/gastrin receptor were determined by Western blot and radioligand binding studies. Amylase secretion and inositol phosphate production assays were used in functional characterization. RESULTS: The CCK-B/gastrin receptor was expressed in the exocrine pancreas and had typical molecular and binding features. CCK and sulfated gastrin stimulated enzyme secretion with identical potencies and efficacies. They activated phospholipase C, but CCK was 60-fold less potent than sulfated gastrin. CONCLUSIONS: The data show that the CCK-B/gastrin receptor mediates exocytosis in acinar cells and can differentially couple to phospholipase C depending on the agonist. The ElasCCKB mice provide a useful model to study phospholipase C-dependent and -independent intracellular transduction pathways leading to pancreatic exocrine secretion.

Amylases↗
PubMed1998

Differential tissular expression of the CCK(A) and CCK(B) gastrin receptor genes during postnatal development in the calf.

Local and temporal expression of CCK(A) and CCK(B)/gastrin receptor genes was studied in the calf with a quantitative Reverse Transcription-Polymerase Chain Reaction (RT-PCR) method. Cerebral cortex, antrum, fundus, gall bladder, pancreas and liver were analyzed in calves at 0, 2, 7, 21, 28 and 150 days of age. Cerebral cortex and pancreas expressed both receptor genes with a ratio between CCK(A) and CCK(B)/gastrin receptor transcripts varying according to the age. Gall bladder and fundus showed an exclusive expression of CCK(A) and CCK(B)/gastrin receptor mRNAs, respectively, with the highest levels of transcripts in newborn and 28-day-old calves. The rank order for CCK(A) receptor mRNA expression was gall bladder > pancreas > cerebral cortex >>> antrum and that for CCK(B)/gastrin receptor mRNA expression was cerebral cortex / pancreas / fundus >> antrum. No CCK(A) and CCK(B)/gastrin receptor mRNA was detected in liver, regardless of the age of calves. The present data represent a basis for a better understanding of the ontogeny of physiological functions linked to the CCK(A) and CCK(B)/gastrin receptors.

Aging↗
PubMed1998

Role of the region 3' to Xist exon 6 in the counting process of X-chromosome inactivation.

During early embryogenesis of female mammals, one of the two X chromosomes is randomly chosen to be inactivated in each cell, leading to the transcriptional silencing of thousands of genes on this chromosome. This random X-inactivation process also occurs during in vitro differentiation of female embryonic stem (ES) cells. A locus on the X chromosome, the X inactivation centre (Xic) is initially 'counted', given that at least two copies of Xic must be present per diploid genome in order for inactivation to occur. The counting process ensures that one X chromosome remains active in diploid cells. In the mouse, the essential functions of Xic can be assured by a 450-kb region containing the Xist gene. Xist maps within Xic (refs 7-10) and is necessary in cis for inactivation. The Xist transcript is a 15-kb RNA which is confined within the nucleus and coats the inactive X chromosome. In order to characterize functional elements within Xic and the Xist gene, we created a 65-kb cre/loxP deletion extending 3' to Xist exon 6. In undifferentiated ES cells, Xist expression from the deleted X chromosome was markedly reduced. In differentiated XX ES cells containing one deleted X chromosome, the X inactivation process still occurred but was never initiated from the unmutated X chromosome. In differentiated ES cells that were essentially XO, the mutated Xic was capable of initiating X inactivation, even in the absence of another Xic. These results demonstrate a role for the region 3' to Xist exon 6 in the counting process and suggest that counting is mediated by a repressive mechanism which prevents inactivation of a single X chromosome in diploid cells.

A Kinase Anchor Proteins↗
PubMed1997

Differential expression of the CCK-A and CCK-B/gastrin receptor genes in human cancers of the esophagus, stomach and colon.

The expression of cholecystokinin (CCK) and gastrin (G) receptors in human gastrointestinal cancers remains poorly documented and is still of a controversial nature. We have measured the levels of mRNA for CCK-A and CCK-B/gastrin receptors using quantitative reverse transcription-polymerase chain reaction (RT-PCR) in primary digestive cancers and hepatic metastases. CCK-A-receptor mRNA was detected in 5 out of 8 esophageal cancers (0.1-1 fg/microg), in 5 out of 8 gastric cancers (0.05-4.2 fg/microg) and in 5 out of 12 colon cancers (0.1-1 fg/microg RNA). CCK-B/gastrin mRNA was not detected in esophageal cancers but was detected in 7 out of 8 gastric cancers (0.05-5.2 fg/microg), and in only 2 out of 12 colon adenocarcinomas (0.05-1 fg/microg RNA). The expression of the CCK-A receptor in esophageal, gastric and colon cancers and of the CCK-B/gastrin receptor in the majority of gastric adenocarcinomas screened may be an important indicator of the influence of CCK and gastrin of local or systemic origin on the growth of these cancers.

Adenocarcinoma↗
PubMed1997

mRNAs encoding CCKB but not CCKA receptors are expressed in human T lymphocytes and Jurkat lymphoblastoid cells.

We previously reported the existence of pharmacologically related gastrin/CCKB type receptors (CCKB-R) in a variant of Jurkat T lymphoblastoid cells (JK(CD3- CD4+)). We studied here the expression of mRNAs encoding CCKA and CCKB receptors in various human white cells by means of Reverse Transcription-Polymerase Chain Reaction (RT-PCR). Using CCKB-R specific primers, we detected a significant expression of CCKB-R mRNA in JK(CD3- CD4+) cells. These transcripts were also expressed, at a lower level, in two other Jurkat clones (JK(CD3+ CD4-) and JK(CD3+ CD4+)), in peripheral blood lymphocytes (PBL) and in purified CD4+ and CD8+ lymphocytes. Activation of Jurkat cells and PBL by T cells mitogenic lectins (jacalin, phytohemaglutinin) did not modify CCKB-R mRNA expression. In all these cells, using CCKA-R specific primers, we could not amplify any specific cDNA fragment corresponding to this receptor. Neither CCKB-R nor CCKA-R mRNAs could be detected in monocytic cells. Our data show for the first time a constitutive expression of CCKB-R transcripts in lymphoid cells. Moreover, the modulation of immunocyte functions by cholecystokinin-related peptides could occur through CCKB-R rather than CCKA-R and affect lymphocytes rather than monocytes.

CD3 Complex↗
PubMed1997

CholecystokininB receptor from human Jurkat lymphoblastic T cells is involved in activator protein-1-responsive gene activation.

The aim of this study was to analyze the role of cholecystokinin (CCK(B)) receptor in human lymphoblastic Jurkat T cells. We investigated the trophic effect resulting from activation of such a receptor by using the reporter gene strategy. For this purpose, we transiently transfected Jurkat T cells with the reporter plasmid p[(TRE)3-tk-Luc] and found that CCK-8 was able to dose-dependently induce luciferase expression related to activator protein-1 (AP-1) activation with a maximal response identical to that obtained with compounds known to activate AP-1 complex (quantitatively, the same level of induction was obtained with 1 nM 12-O-tetradecanoylphorbol-13-acetate, 100 microM diacylglycerol, or 4 nM epidermal growth factor). The involvement of the CCK(B) receptor in such a stimulation was demonstrated by the inhibiting effect of the selective CCK(B) receptor antagonist PD-135,158. This effect was confirmed in COS-7 cells transfected with the cDNA of CCK(B) receptor cloned from Jurkat T cells. To better understand the AP-1-dependent luciferase expression in Jurkat T cells, we tested two specific inhibitors of serine/threonine phosphatases-1 and -2A: okadaic acid and calyculin A. These compounds strongly increased the phorbol-12-myristate-13-acetate response, whereas we have not observed a contribution of phosphatase inhibitors on a CCK-8-induced luciferase activity. To confirm that CCK(B) receptors are involved in AP-1 response, we investigated the CCK-8 effect on interleukin-2 expression, a natural endogenous gene regulated by several factors, including AP-1. In Jurkat T cells activated by phorbol-12-myristate-13-acetate and phytohemagglutinin, CCK-8 induced IL-2 expression. This induction was abolished by PD-135,158. Our results indicate that CCK-8 exerts a trophic effect in Jurkat T cells through stimulation of CCK(B) receptors by modulation of expression of AP-1-regulated genes.

Animals↗
PubMed1997

X-chromosome inactivation in mammals.

The inactive X chromosome differs from the active X in a number of ways; some of these, such as allocyclic replication and altered histone acetylation, are associated with all types of epigenetic silencing, whereas others, such as DNA methylation, are of more restricted use. These features are acquired progressively by the inactive X after onset of initiation. Initiation of X-inactivation is controlled by the X-inactivation center (Xic) and influenced by the X chromosome controlling element (Xce), which causes primary nonrandom X-inactivation. Other examples of nonrandom X-inactivation are also presented in this review. The definition of a major role for Xist, a noncoding RNA, in X-inactivation has enabled investigation of the mechanism leading to establishment of the heterochromatinized X-chromosome and also of the interactions between X-inactivation and imprinting as well as between X-inactivation and developmental processes in the early embryo.

Animals↗
PubMed1997

Pancreatic cancer cell regulation by lipids and by basic fibroblast growth factor expression.

High fat intake is a risk factor for pancreatic cancer. Lipids may act either directly or in cooperation with growth-promoting polypeptides. In this study, the role of serum lipids, and mainly the often expressed intracellular basic fibroblast growth factor (bFGF) isoforms in cancer cells, was analyzed in pancreatic tumor cell proliferation. Serum lipids alone induced a 1.9-fold increase of human pancreatic cancer cell growth (p < 0.001). Treatment with bFGF had a weak mitogenic effect (1.2- to 1.3-fold increase) compared with those of insulin and transferrin (1.7- to 1.6-fold increase, respectively). The bFGF expression by a rat pancreatic cancer cell line that was transfected with bFGF cDNAs modified cell lipid contents and induced a higher proliferation rate than that found with the exogenous bFGF. Combined extra- and intracellular bFGFs increased cell growth by two to three times (p < 0.001), regardless the presence of extracellular lipids. The results obtained reflect the direct mitogenic effect of serum lipids and suggest that the endogenous bFGF of high molecular weight may be implicated in pancreatic cancer cell growth. By modifying cell lipids, bFGFs may interfere with other cell functions, like signal transduction.

Animals↗
PubMed1996

Molecular cloning, developmental expression and pharmacological characterization of the CCKB/gastrin receptor in the calf pancreas.

We have cloned the calf predominant pancreatic cholecystokinin B (CCKB)/gastrin receptor cDNA. It encodes a 454 amino acid protein with 90% identity with the CCKB/gastrin receptor cloned in other species and tissues. However, the calf pancreatic CCKB/gastrin receptor contains a pentapeptide cassette within the third intracellular loop which is absent in the cloned human brain and stomach receptor. Quantification of the CCKB/gastrin receptor mRNA levels by reverse transcription polymerase chain reaction demonstrated the same level of transcripts at birth, +7 and +28 days. On the other hand, binding study with pancreatic membranes showing a dramatic increase (600-fold) in the number of CCKB/gastrin receptor sites between at birth and +28 days indicates that the development of the calf pancreatic CCKB/gastrin receptor occurs during the first 4 weeks of post-natal life. COS monkey cells (COS-7 cells) transiently transfected by the cloned cDNA exhibit binding of 125I-Bolton-Hunter-[Thr28,Ahx31]CCK-(25-33) and 125I-Bolton-Hunter-[Leu15]human gastrin-(2-17) to two affinity classes of sites. Kd values of the high affinity binding components indicate a 4-fold higher affinity of the receptor for sulfated gastrin than for CCK. Finally, the recombinant receptor is coupled to G proteins and [Ca2+]i mobilization, and is expressed as a glycoprotein of 82 kDa.

Amino Acid Sequence↗
PubMed1996

A 94 kb genomic sequence 3' to the murine Xist gene reveals an AT rich region containing a new testis specific gene Tsx.

X chromosome inactivation in both mouse and human requires the presence of a cis acting locus, the X inactivation centre. This locus is thought to be involved in the initiation and spreading of the inactivation signal in early development. In order to increase our understanding of the mouse X inactivation centre, a 94 kb region immediately distal to the Xist gene has been sequenced and analysed for the presence of transcription units and/or potential cis acting regulatory elements. We have identified a novel gene, Tsx, lying 40 kb 3' from Xist. Tsx is expressed specifically in the testis and shows no convincing homology to proteins currently in the databases. A rat homologue, also X linked, has been isolated. The mouse and rat Tsx sequences are highly divergent, suggesting that part of the X inactivation centre, including both Xist and Tsx are subject to relatively weak evolutionary constraints.

Animals↗
PubMed1995

Identification of a region of the N-terminal of the human CCKA receptor essential for the high affinity interaction with agonist CCK.

The discovery of an N-terminally truncated isoform of the cholecystokinin A subtype (CCKA) receptor exhibiting an atypical pharmacology led us to study the effects of N-terminal truncation on the pharmacology of the human CCKA receptor. We cloned the cDNA encoding the full CCKA receptor and constructed two truncated forms, one which lacked the first 37 amino acids (CCKAT38) and another which lacked the first 42 amino acids (CCKAT43). Expression of the receptors in COS-7 cells showed that the CCKAT38 receptor displayed a pharmacological profile identical to that of the full receptor. In contrast, the CCKAT43 receptor did not directly bind agonist CCK9; however, the agonist could compete for binding at low affinity sites. Binding of the partial agonist JMV180 and the antagonist JMV179 were unaffected. These results identify for the first time a part of the N-terminal, close to the membrane, of the human CCKA receptor that is essential for the high affinity interaction with CCK.

Amino Acid Sequence↗