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1.
Neurobiol Dis ; 69: 93-107, 2014 Sep.
Article in English | MEDLINE | ID: mdl-24878511

ABSTRACT

After incomplete spinal cord injury (SCI), patients and animals may exhibit some spontaneous functional recovery which can be partly attributed to remodeling of injured neural circuitry. This post-lesion plasticity implies spinal remodeling but increasing evidences suggest that supraspinal structures contribute also to the functional recovery. Here we tested the hypothesis that partial SCI may activate cell-signaling pathway(s) at the supraspinal level and that this molecular response may contribute to spontaneous recovery. With this aim, we used a rat model of partial cervical hemisection which injures the bulbospinal respiratory tract originating from the medulla oblongata of the brainstem but leads to a time-dependent spontaneous functional recovery of the paralyzed hemidiaphragm. We first demonstrate that after SCI the PI3K/Akt signaling pathway is activated in the medulla oblongata of the brainstem, resulting in an inactivation of its pro-apoptotic downstream target, forkhead transcription factor (FKHR/FOXO1A). Retrograde labeling of medullary premotoneurons including respiratory ones which project to phrenic motoneurons reveals an increased FKHR phosphorylation in their cell bodies together with an unchanged cell number. Medulla infusion of the PI3K inhibitor, LY294002, prevents the SCI-induced Akt and FKHR phosphorylations and activates one of its death-promoting downstream targets, Fas ligand. Quantitative EMG analyses of diaphragmatic contractility demonstrate that the inhibition of medulla PI3K/Akt signaling prevents spontaneous respiratory recovery normally observed after partial cervical SCI. Such inhibition does not however affect either baseline contractile frequency or the ventilatory reactivity under acute respiratory challenge. Together, these findings provide novel evidence of supraspinal cellular contribution to the spontaneous respiratory recovery after partial SCI.


Subject(s)
Forkhead Transcription Factors/metabolism , Medulla Oblongata/physiopathology , Nerve Tissue Proteins/metabolism , Oncogene Protein v-akt/metabolism , Recovery of Function/physiology , Respiration , Spinal Cord Injuries/physiopathology , Animals , Cervical Vertebrae , Diaphragm/physiopathology , Disease Models, Animal , Female , Functional Laterality , Medulla Oblongata/drug effects , Medulla Oblongata/pathology , Motor Neurons/pathology , Motor Neurons/physiology , Neuronal Plasticity/drug effects , Neuronal Plasticity/physiology , Oncogene Protein v-akt/antagonists & inhibitors , Phosphatidylinositol 3-Kinases/metabolism , Phosphoinositide-3 Kinase Inhibitors , Phrenic Nerve/pathology , Phrenic Nerve/physiopathology , Rats, Sprague-Dawley , Recovery of Function/drug effects , Respiration/drug effects , Signal Transduction/drug effects , Spinal Cord Injuries/pathology , Time Factors
2.
Front Horm Res ; 42: 93-106, 2014.
Article in English | MEDLINE | ID: mdl-24732928

ABSTRACT

Prader-Willi syndrome (PWS), first described in 1956, is considered as a paradigm of a neurodevelopmental disorder with severe and early obesity with hyperphagia and impaired satiety. The improved knowledge in the natural history and recent data on genetics offer new perspectives for understanding the metabolic and endocrine dysfunctions and possibly for treatment. Natural history of the disease has been described due to the early diagnosis performed in the first months of life and various nutritional phases have been described. In addition, there is clear evidence that the abnormal feeding behavior is included in the behavioral problems. Brain imaging studies have shown that some brain regions may be important in PWS. The role of SNORD116 gene cluster is detailed and its links with circadian rhythm and brain and hypothalamus development. Pathophysiology of the abnormal ghrelin levels and of OT dysfunction is documented. While no effect on appetite and weight regulation has been reported with ghrelin antagonists, OT has been shown to improve some of the behavioral problems in adults. We discuss our hypothesis of an abnormal ghrelin/OT/dopamine pathway which may explain the switch of nutritional phases and behavior. These new aspects offer an opportunity for therapeutic use and possible early intervention.


Subject(s)
Brain/metabolism , Dopamine/metabolism , Ghrelin/metabolism , Hyperphagia/metabolism , Prader-Willi Syndrome/metabolism , Feeding Behavior , Humans
3.
Cell Death Differ ; 15(12): 1921-9, 2008 Dec.
Article in English | MEDLINE | ID: mdl-18772898

ABSTRACT

NRAGE (also known as Maged1, Dlxin) is a member of the MAGE gene family that may play a role in the neuronal apoptosis that is regulated by the p75 neurotrophin receptor (p75NTR). To test this hypothesis in vivo, we generated NRAGE knockout mice and found that NRAGE deletion caused a defect in developmental apoptosis of sympathetic neurons of the superior cervical ganglia, similar to that observed in p75NTR knockout mice. Primary sympathetic neurons derived from NRAGE knockout mice were resistant to apoptosis induced by brain-derived neurotrophic factor (BDNF), a pro-apoptotic p75NTR ligand, and NRAGE-deficient sympathetic neurons show attenuated BDNF-dependent JNK activation. Hair follicle catagen is an apoptosis-like process that is dependent on p75NTR signaling; we show that NRAGE and p75NTR show regulated co-expression in the hair follicle and that identical defects in hair follicle catagen are present in NRAGE and p75NTR knockout mice. Interestingly, NRAGE knockout mice have severe defects in motoneuron apoptosis that are not observed in p75NTR knockout animals, raising the possibility that NRAGE may facilitate apoptosis induced by receptors other than p75NTR. Together, these studies demonstrate that NRAGE plays an important role in apoptotic-signaling in vivo.


Subject(s)
Adaptor Proteins, Signal Transducing/metabolism , Apoptosis , Neoplasm Proteins/metabolism , Receptor, Nerve Growth Factor/metabolism , Animals , Fertility , Gene Targeting , Hair Follicle/growth & development , Hair Follicle/pathology , Mice , Mice, Knockout , Motor Neurons/cytology , Mutation/genetics , Neoplasm Proteins/deficiency , Sympathetic Nervous System/cytology , Trigeminal Ganglion/abnormalities
4.
J Intellect Disabil Res ; 52(Pt 5): 426-36, 2008 May.
Article in English | MEDLINE | ID: mdl-18298478

ABSTRACT

Introduction The present study reports cross-cultural comparisons of body mass index (BMI) and growth in Prader-Willi syndrome, a neurodevelopmental disorder associated with obesity, growth restriction and mild learning disability. Our objectives were to: (1) compare rates of obesity in adults with Prader-Willi syndrome (PWS) in France, with data available from Belgium, the UK and the USA; (2) compare growth of French children with PWS with their counterparts in Germany and the USA; and (3) evaluate the contribution of genetic, medical and social parameters to obesity outcome in French children and adults with PWS. Method (1) Cross-sectional comparison of BMI of 40 French adults, 38 Belgian adults, 46 British adults and 292 North American adults; (2) Construction of growth curves for French children aged 2-20 years from longitudinal data for 150 individuals with PWS, and comparison with published growth curves from Germany and the USA; and (3) Longitudinal regression analysis of 141 French children and adults to determine the factors contributing to obesity outcome. Results A total of 82.5% French adults with PWS have BMI > 30 compared with 65.8% in Belgium (n.s.), 58.2% in the USA (P < 0.005), and 54.3% in the UK (P < 0.01). Higher rates of obesity in females vs. males were found in the USA sample (P < 0.001) but not in the other samples. In contrast to adults, growth curves for French children with PWS show similar rates of growth compared with children with PWS in Germany and the USA. The principal determining factors of BMI status in the French PWS population are age (P < 0.0001), cohort (born within the last 15 years vs. born over 15 years ago, P < 0.0002) and growth hormone replacement therapy (P < 0.0002). Significant subsidiary effects include domestic situation (P < 0.0001), genetic diagnosis (P < 0.0001) and age of diagnosis (P < 0.0001). Conclusions French adults with PWS have significantly higher rates of obesity than adults in the UK and the USA, but growth in French children with PWS is similar to the USA and Germany. Clinical management has a greater impact on obesity outcome in PWS than cultural factors.


Subject(s)
Cross-Cultural Comparison , Developmental Disabilities/epidemiology , Obesity/epidemiology , Prader-Willi Syndrome/epidemiology , Adolescent , Adult , Age Distribution , Belgium/epidemiology , Body Mass Index , Child , Child, Preschool , Cohort Studies , Comorbidity , Cross-Sectional Studies , Female , France , Genetic Predisposition to Disease , Germany/epidemiology , Humans , Longitudinal Studies , Male , Middle Aged , Sex Distribution , Socioeconomic Factors , United Kingdom/epidemiology , United States/epidemiology
5.
Early Hum Dev ; 83(7): 471-8, 2007 Jul.
Article in English | MEDLINE | ID: mdl-17055193

ABSTRACT

BACKGROUND: Imprinted genes are considered to play an important role in growth and early development but much of the research is based on animal studies. AIM: This study reports clinical data from a French population concerning prenatal, perinatal and postnatal complications in Prader-Willi syndrome (PWS), a genetically imprinted neurodevelopmental disorder associated with growth retardation, intellectual impairment and obesity. STUDY DESIGN: Data from family health records concerning prenatal, perinatal and postnatal complications were collected from 52 French people with the deletion form (DEL), and 34 French people with the maternal disomy form of PWS (UPD) and compared against national norms and between groups. RESULTS: Significant findings include: a history of miscarriage, high rate of polyhydramnios (12/34 UPD, 11/52 DEL), a high rate of induced labour, a high rate of Caesarian section (20/34 UPD, 26/52 DEL), small gestational age (10/34 UPD, 22/52 DEL), hypotonia (34/34 UPD, 49/52 DEL), and suckling deficit (25/34 UPD, 46/52 DEL). Significant differences between genetic subtypes include a higher rate of induced labour in UPD (27/34 UPD, 25/52 DEL), an increased risk of premature term in UPD (9/34 UPD vs. 4/52 DEL), raised maternal age in UPD (36.4 years vs. 29.3 years), low birth weight for newborns with a deletion form of PWS (girls 2.8 kg, boys 2.7 kg), a positive correlation between parental weight and offspring birth weight only for patients with UPD (UPD maternal: r=0.62, paternal: r=0.51). CONCLUSION: The results indicate significant intrauterine disturbance in PWS, particularly in PWS due to UPD, but a more significant weight disturbance for PWS due to deletion.


Subject(s)
Fetal Growth Retardation/etiology , Genomic Imprinting , Prader-Willi Syndrome/complications , Adolescent , Adult , Child , Child, Preschool , Chromosome Aberrations , DNA/analysis , Female , Fetal Growth Retardation/diagnosis , Humans , Infant , Infant, Newborn , Infant, Newborn, Diseases/etiology , Male , Middle Aged , Prader-Willi Syndrome/genetics , Pregnancy , Pregnancy Complications
6.
Eur J Hum Genet ; 9(7): 519-26, 2001 Jul.
Article in English | MEDLINE | ID: mdl-11464243

ABSTRACT

Imprinting defects in 15q11-q13 are a rare but significant cause of Prader-Willi syndrome (PWS) and Angelman syndrome (AS). Patients with an imprinting defect have apparently normal chromosomes 15 of biparental origin, but are recognised by @parental DNA methylation at D15S63 (PW71) or SNURF-SNRPN exon 1. We have investigated the methylation status of five additional loci in 12 such patients with or without a deletion in the imprinting centre. In each patient, the imprinting defect affected all loci tested. During routine diagnostic testing we identified four patients who had a normal methylation pattern at SNURF-SNRPN exon 1, but an abnormal pattern at D15S63. In two of these patients, who were suspected of having PWS, this change was restricted to D15S63. In two patients suspected of having AS, several but not all loci were affected. Using a newly developed methylation-specific PCR test for D15S63 we found that these methylation changes are rare in patients suspected of having AS. Although we can not prove that the methylation changes in the four patients are causally related to their disease, our findings demonstrate that spatially restricted changes in methylation can occur. In some cases, these changes may reflect incomplete imprint spreading.


Subject(s)
Angelman Syndrome/genetics , Chromosomes, Human, Pair 15/genetics , DNA Methylation , Prader-Willi Syndrome/genetics , Blotting, Southern , DNA/genetics , DNA/metabolism , Family Health , Female , Genomic Imprinting , Humans , Male , Microsatellite Repeats
7.
Hum Mol Genet ; 9(20): 3101-10, 2000 Dec 12.
Article in English | MEDLINE | ID: mdl-11115855

ABSTRACT

Prader-Willi syndrome (PWS) is a complex neurogenetic disorder with considerable clinical variability that is thought in large part to be the result of a hypothalamic defect. PWS results from the absence of paternal expression of imprinted genes localized in the 15q11-q13 region; however, none of the characterized genes has so far been shown to be involved in the etiology of PWS. Here, we provide a detailed investigation of a mouse model deficient for NECDIN: Linked to the mutation, a neonatal lethality of variable penetrance is observed. Viable NECDIN: mutants show a reduction in both oxytocin-producing and luteinizing hormone-releasing hormone (LHRH)-producing neurons in hypothalamus. This represents the first evidence of a hypothalamic deficiency in a mouse model of PWS. NECDIN:-deficient mice also display increased skin scraping activity in the open field test and improved spatial learning and memory in the Morris water maze. The latter features are reminiscent of the skin picking and improved spatial memory that are characteristics of the PWS phenotype. These striking parallels in hypothalamic structure, emotional and cognitive-related behaviors strongly suggest that NECDIN is responsible for at least a subset of the multiple clinical manifestations of PWS.


Subject(s)
Nerve Tissue Proteins/genetics , Neurons/physiology , Nuclear Proteins/genetics , Prader-Willi Syndrome/genetics , Animals , Cognition/physiology , Cognition Disorders/genetics , Disease Models, Animal , Gonadotropin-Releasing Hormone/metabolism , Hypothalamus/metabolism , Hypothalamus/physiology , Mice , Mice, Knockout , Nerve Tissue Proteins/deficiency , Nerve Tissue Proteins/physiology , Nuclear Proteins/deficiency , Nuclear Proteins/physiology , Oxytocin/metabolism , Prader-Willi Syndrome/psychology , Psychomotor Performance/physiology
8.
Hum Mol Genet ; 8(13): 2497-505, 1999 Dec.
Article in English | MEDLINE | ID: mdl-10556298

ABSTRACT

Prader-Willi syndrome (PWS) is a complex neurogenetic disorder. The phenotype is likely to be a contiguous gene syndrome involving genes which are paternally expressed only, located in the human 15q11-q13 region. Four mouse models of PWS have been reported but these do not definitively allow the delineation of the critical region and the associated genes involved in the aetiology of PWS. Moreover, targeted mutagenesis of mouse homologues of the human candidate PWS genes does not appear to result in any of the features of PWS. Therefore, the isolation of new genes in this region remains crucial for a better understanding of the molecular basis of PWS. In this manuscript, we report the characterization of MAGEL2 and its mouse homologue Magel2. These are located in the human 15q11-q13 and mouse 7C regions, in close proximity to NDN / Ndn. By northern blot analysis we did not detect any expression of MAGEL2 / Magel2 but by RT-PCR analysis, specific expression was detected in fetal and adult brain and in placenta. Both genes are intronless with tandem direct repeat sequences contained within a CpG island in the 5'-untranscribed region. The transcripts encode putative proteins that are homologous to the MAGE proteins and NDN. Moreover, MAGEL2 / Magel2 are expressed only from the paternal allele in brain, suggesting a potential role in the aetiology of PWS and its mouse model, respectively.


Subject(s)
Chromosome Mapping , Prader-Willi Syndrome/genetics , Proteins/genetics , 5' Untranslated Regions , Amino Acid Sequence , Animals , Antigens, Neoplasm , Base Sequence , Brain/metabolism , Chromosomes, Human, Pair 15 , CpG Islands , Gene Expression , Genomic Imprinting , Humans , Mice , Mice, Inbred Strains , Molecular Sequence Data , Pedigree , Proteins/metabolism , Reverse Transcriptase Polymerase Chain Reaction , Sequence Alignment , Tandem Repeat Sequences
9.
Eur J Hum Genet ; 5(5): 324-32, 1997.
Article in English | MEDLINE | ID: mdl-9412790

ABSTRACT

Prader-Willi syndrome (PWS) is a neurogenetic disorder resulting from the loss of paternal expression of gene(s) localized in the 15q11-q12 region. A new human gene encoding a putative protein with high homology to the mouse NECDIN protein has recently been characterized and mapped to chromosome 15q11-q12. It is expressed from the paternal allele only, suggesting its potential involvement in PWS. We now report the localization of the mouse Necdin gene in a region of conserved synteny to the human PWS region. We demonstrate the paternal specific expression of Necdin in the mouse central nervous system, and show that parental alleles display a differential methylation profile in the coding region. Finally, fluorescence in situ hybridization analysis reveals an asynchronous pattern of replication at the Necdin locus. These results clearly demonstrate imprinting of the mouse Necdin gene. Mouse models will be powerful tools in the study of human PWS phenotype and imprinting mechanisms.


Subject(s)
Chromosome Mapping , Gene Expression Regulation, Developmental/genetics , Genomic Imprinting/genetics , Nerve Tissue Proteins/genetics , Nuclear Proteins/genetics , Prader-Willi Syndrome/genetics , Alleles , Animals , Base Sequence , Brain , Brain Chemistry , Conserved Sequence/genetics , Crosses, Genetic , DNA Methylation , DNA Replication , Female , Fetus , Humans , Male , Mice , Mice, Inbred C57BL , Molecular Sequence Data , Muridae , RNA, Messenger/analysis
10.
Nat Genet ; 17(3): 357-61, 1997 Nov.
Article in English | MEDLINE | ID: mdl-9354807

ABSTRACT

Prader-Willi syndrome (PWS) is a neurogenetic disorder that results from the absence of a normal paternal contribution to the 15q11-13 region. The clinical manifestations of PWS are a transient severe hypotonia in the newborn period, with mental retardation, hypogonadism and obesity observed later in development. Five transcripts with exclusive expression from the paternal allele have been isolated, but none of these has been shown to be involved in PWS. In this study, we report the isolation and characterization of NDN, a new human imprinted gene. NDN is exclusively expressed from the paternal allele in the tissues analysed and is located in the PWS region. It encodes a putative protein homologous to the mouse brain-specific NECDIN protein, NDN; as in mouse, expression in brain is restricted to post-mitotic neurons. NDN displays several characteristics of an imprinted locus, including allelic DNA methylation and asynchronous DNA replication. A complete lack of NDN expression in PWS brain and fibroblasts indicates that the gene is expressed exclusively from the paternal allele in these tissues and suggests a possible role of this new gene in PWS.


Subject(s)
Gene Expression Regulation, Developmental , Genomic Imprinting , Nerve Tissue Proteins/genetics , Nuclear Proteins/genetics , Prader-Willi Syndrome/genetics , Angelman Syndrome/genetics , Animals , Blotting, Northern , Chromosome Mapping , Chromosomes, Human, Pair 15 , DNA Methylation , Deoxyribonucleases, Type II Site-Specific/genetics , Female , Humans , In Situ Hybridization/methods , In Situ Hybridization, Fluorescence , Male , Mice , Molecular Sequence Data , Nerve Tissue Proteins/metabolism , Nervous System Physiological Phenomena , Nuclear Proteins/metabolism , Tissue Distribution
11.
Genomics ; 46(3): 397-408, 1997 Dec 15.
Article in English | MEDLINE | ID: mdl-9441743

ABSTRACT

Genes of the MAGE family direct the expression of tumor antigens recognized on a human melanoma by autologous cytolytic T lymphocytes. Twelve closely related MAGE genes are located in the Xq28 region. These genes share 60-98% nucleotide identity in their coding region. The presence of homologous genes in a region of Xp21.3 has been reported previously. We obtained the complete sequence of a 42-kb stretch of this region. It contains four MAGE-related genes, which we propose to name MAGE-B1, B2, B3, and B4 (HGMW-approved symbols MAGEB1, MAGEB2, MAGEB3, and MAGEB4). The coding regions of these genes share 66-81% nucleotide identity and show 45-63% identity with those of the MAGE genes located in Xq28. Like the MAGE genes located in Xq28, the MAGE-B genes are silent in normal tissues with the exception of testis. Like MAGE-1, 2, 3, 4, 6 and 12 (HGMW-approved symbols MAGEA1, 2, 3, 4, 6, and 12), genes MAGE-B1 and MAGE-B2 are expressed in a significant fraction of tumors of various histological types. The transcription of MAGE-B1 and MAGE-B2 can be induced by 5-aza-2'-deoxycytidine, suggesting that the activation of these genes in tumors results from a demethylation process.


Subject(s)
Neoplasm Proteins/genetics , Neoplasms/genetics , X Chromosome , Alternative Splicing , Amino Acid Sequence , Azacitidine/analogs & derivatives , Azacitidine/pharmacology , Base Sequence , DNA, Complementary , Decitabine , Exons , Gene Expression , Humans , Molecular Sequence Data , Sequence Homology, Amino Acid , Sequence Homology, Nucleic Acid , Tumor Cells, Cultured
12.
Am J Hum Genet ; 58(6): 1205-11, 1996 Jun.
Article in English | MEDLINE | ID: mdl-8651297

ABSTRACT

We demonstrate that isolated glycerol kinase (GK) deficiency in three families results from mutation of the Xp21 GK gene. GK mutations were detected in four patients with widely differing phenotypes. Patient 1 had a splice-site mutation causing premature termination. His general health was good despite absent GK activity, indicating that isolated GK deficiency can be silent. Patient 2 had GK deficiency and a severe phenotype involving psychomotor retardation and growth delay, bone dysplasia, and seizures, similar to the severe phenotype of one of the first described cases of GK deficiency. His younger brother, patient 3, also had GK deficiency, but so far his development has been normal. GK exon 17 was deleted in both brothers, implicating additional factors in causation of the severe phenotype of patient 2. Patient 4 had both GK deficiency with mental retardation and a GK missense mutation (D440V). Possible explanations for the phenotypic variation of these four patients include ascertainment bias; metabolic or environmental stress as a precipitating factor in revealing GK-related changes, as has previously been described in juvenile GK deficiency; and interactions with functional polymorphisms in other genes that alter the effect of GK deficiency on normal development.


Subject(s)
Abnormalities, Multiple/genetics , Glycerol Kinase/deficiency , Glycerol Kinase/genetics , Mutation , X Chromosome , Abnormalities, Multiple/enzymology , Adolescent , Alternative Splicing , Base Sequence , Child , Child, Preschool , Chromosome Mapping , DNA Primers , Exons , Fibroblasts/enzymology , Humans , Introns , Leukocytes/enzymology , Male , Middle Aged , Molecular Sequence Data , Phenotype , Polymerase Chain Reaction , Restriction Mapping , Sequence Deletion
13.
Proc Natl Acad Sci U S A ; 92(11): 4987-91, 1995 May 23.
Article in English | MEDLINE | ID: mdl-7761436

ABSTRACT

A human gene with strong homology to the MAGE gene family located in Xq27-qter has been isolated by using exon-trapping of cosmids in the Xp21.3 region. We have mapped and sequenced cDNA and genomic clones corresponding to this gene, MAGE-Xp, and shown that the last exon contains the open reading frame and is present in a minimum of five copies in a 30-kb interval. MAGE-Xp is expressed only in testis and, unlike the Xq27-qter MAGE genes, it is not expressed in any of 12 different tumor tissues tested. However, the gene and predicted protein structure are conserved, suggesting a similar function. MAGE-Xp is located in the 160-kb critical interval defined for the locus involved in sex determination within Xp21 and is 50 kb distal to the DAX-1 gene, which is responsible for X-chromosome-linked adrenal hypoplasia congenita.


Subject(s)
Hominidae/genetics , Multigene Family , Neoplasm Proteins , Proteins/genetics , X Chromosome , Adrenal Glands/abnormalities , Amino Acid Sequence , Animals , Antigens, Neoplasm , Base Sequence , Chromosome Mapping , Chromosomes, Artificial, Yeast , Conserved Sequence , Cosmids , DNA Primers , DNA, Complementary , Exons , Gene Library , Genetic Linkage , Humans , Hypogonadism/genetics , Introns , Male , Molecular Sequence Data , Neoplasms/genetics , Neoplasms/metabolism , Oligonucleotide Probes , Polymerase Chain Reaction , Protein Biosynthesis , Restriction Mapping , Sequence Homology, Amino Acid , Testis/metabolism
14.
Oncogene ; 9(12): 3683-8, 1994 Dec.
Article in English | MEDLINE | ID: mdl-7970727

ABSTRACT

The Bmx sequence was identified and cloned during our search for novel tyrosine kinase genes expressed in human bone marrow cells. Bmx cDNA comprises a long open reading frame of 675 amino acids, containing one SH3, one SH2 and one tyrosine kinase domain, which are about 70% identical with Btk, Itk and Tec and somewhat less with Txk tyrosine kinase sequences. The amino terminal sequences of these four tyrosine kinases are about 40% identical and each contains a so-called pleckstrin homology domain. The 2.7 kb Bmx mRNA was expressed in endothelial cells and several human tissues by Northern blotting and an 80 kD Bmx polypeptide was detected in human endothelial cells. Immunoprecipitates of COS cells transfected with a Bmx expression vector and NIH3T3 cells expressing a Bmx retrovirus contained a tyrosyl phosphorylated Bmx polypeptide of similar molecular weight. The BMX gene was located in chromosomal band Xp22.2 between the DXS197 and DXS207 loci. Interestingly, chromosome X also contains the closest relative of BMX, the BTK gene, implicated in X-linked agammaglobulinemia. The BMX gene thus encodes a novel nonreceptor tyrosine kinase, which may play a role in the growth and differentiation of hematopoietic cells.


Subject(s)
Protein-Tyrosine Kinases/genetics , X Chromosome , 3T3 Cells , Amino Acid Sequence , Animals , Cell Line , Chromosome Mapping , Cloning, Molecular , DNA, Complementary , Humans , Mice , Molecular Sequence Data , Multigene Family , Sequence Homology, Amino Acid
15.
Nature ; 372(6507): 635-41, 1994 Dec 15.
Article in English | MEDLINE | ID: mdl-7990953

ABSTRACT

X-linked adrenal hypoplasia congenita is a developmental disorder of the human adrenal gland that results in profound hormonal deficiencies and is lethal if untreated. We have isolated the gene responsible for the disease, DAX-1, which is deleted or mutated in X-linked adrenal hypoplasia patients. DAX-1 encodes a new member of the nuclear hormone receptor superfamily displaying a novel DNA-binding domain. The DAX-1 product acts as a dominant negative regulator of transcription mediated by the retinoic acid receptor.


Subject(s)
Adrenal Insufficiency/genetics , DNA-Binding Proteins/genetics , Mutation , Receptors, Cytoplasmic and Nuclear/genetics , Receptors, Retinoic Acid/genetics , Repressor Proteins , Transcription Factors/genetics , X Chromosome , Adrenal Insufficiency/congenital , Amino Acid Sequence , Animals , Base Sequence , Cell Nucleus/metabolism , Chromosome Mapping , DAX-1 Orphan Nuclear Receptor , DNA, Complementary/metabolism , DNA-Binding Proteins/metabolism , Female , Gene Deletion , Gene Expression , Genetic Linkage , Humans , Male , Molecular Sequence Data , Receptors, Cytoplasmic and Nuclear/metabolism , Receptors, Retinoic Acid/metabolism , Sequence Homology, Amino Acid , Sex Differentiation/genetics , Transcription, Genetic
16.
Nature ; 372(6507): 672-6, 1994 Dec 15.
Article in English | MEDLINE | ID: mdl-7990958

ABSTRACT

Adrenal hypoplasia congenita (AHC) is an X-linked disorder characterized by primary adrenal insufficiency. Hypogonadotropic hypogonadism (HHG) is frequently associated with this disorder but is thought not to be caused by the low adrenal androgen levels due to adrenal hypoplasia. It is uncertain whether there are two distinct yet physically linked genes responsible for AHC and HHG or a single gene responsible for both diseases. AHC can occur as a part of a contiguous deletion syndrome together with Duchenne muscular dystrophy (DMD) and/or glycerol kinase deficiency (GKD). From the analysis of deletions, the following gene order has been deduced: Xpter-AHC-GKD-DMD-cen. An AHC critical region of 200-500 kilobases has been defined by physical mapping and partially overlaps with a 160-kilobase dosage-sensitive sex (DSS) reversal critical region. The DAX-1 (DSS-AHC critical region on the X, gene 1) gene was isolated and found to encode a new member of the nuclear hormone receptor family. Here we report that DAX-1 is deleted in 14 patients and point mutations were found in the coding region in DNA from 12 unrelated individuals. All AHC patients over 14 years old and with only point mutations in DAX-1 were also diagnosed with HHG, confirming that the DAX-1 gene is responsible for both X-linked AHC and HHG. But in four sporadic cases and a single familial case, no point mutations were found, suggesting genetic heterogeneity or differential expression of DAX-1.


Subject(s)
Adrenal Insufficiency/congenital , Adrenal Insufficiency/genetics , DNA-Binding Proteins/genetics , Hypogonadism/genetics , Point Mutation , Receptors, Cytoplasmic and Nuclear/genetics , Receptors, Retinoic Acid/genetics , Repressor Proteins , Transcription Factors/genetics , X Chromosome , Base Sequence , Child , Child, Preschool , DAX-1 Orphan Nuclear Receptor , DNA , Genetic Linkage , Humans , Infant , Infant, Newborn , Molecular Sequence Data , Restriction Mapping
17.
Clin Endocrinol (Oxf) ; 40(6): 807-8, 1994 Jun.
Article in English | MEDLINE | ID: mdl-8033374

ABSTRACT

We report a patient with X-linked adrenal hypoplasia congenita and hypogonadotrophic hypogonadism in whom there were no clinical or biochemical features of either glycerol kinase deficiency or Duchenne muscular dystrophy. (The adrenal hypoplasia congenita and glycerol kinase loci map in Xp21 distal to Duchenne muscular dystrophy, and proximal to DXS727). DNA isolated from our patient was analysed by PCR amplification with primers for appropriate loci in the Xp21 region. This analysis revealed the absence of DXS319, which lies near the adrenal hypoplasia congenita deletion critical region, and the presence of DXS727, which is distal to the gene. The absence of glycerol kinase deficiency biochemically and clinically was consistent with the presence of one glycerol kinase exon product from PCR primers P17/P18 which lies within the glycerol kinase gene. The hypogonadotrophic hypogonadism is universally found in X-linked adrenal hypoplasia congenita and is thought to be pituitary in origin. These findings suggest that a gene locus resulting in hypogonadotrophic hypogonadism is present in the Xp21 region and is an integral part of the adrenal hypoplasia congenita gene or in close relationship to it.


Subject(s)
Adrenal Glands/pathology , Gene Deletion , Genetic Linkage , Hypogonadism/congenital , Hypogonadism/genetics , X Chromosome , Adolescent , Humans , Hypogonadism/complications , Male , Polymerase Chain Reaction
18.
Am J Trop Med Hyg ; 49(3): 364-9, 1993 Sep.
Article in English | MEDLINE | ID: mdl-8372958

ABSTRACT

To construct a DNA probe specific for protozoa that cause visceral leishmaniasis, we cloned Pst I fragments of Leishmania infantum genomic DNA into a Bluescript II SK vector. A clone of 4.3 kb that contained a highly repetitive sequence was isolated and cut with three restriction enzymes: Hae III, Rsa I, and Sau 3A. After a new molecular cloning step, we isolated and sequenced a 140-basepair (bp) fragment. Two oligonucleotides were synthesized to be used as primers for a polymerase chain reaction. Using this probe, we detected an amount of DNA equivalent to one promastigote of L. infantum. This probe showed a high specificity; all protozoa tested that cause visceral leishmaniasis and L. major (one of the causative agents of Old World cutaneous leishmaniasis) showed a 100-bp amplified sequence, whereas other Leishmania strains showed a signal of a different size or else no signal. Moreover, no amplified sequence was obtained with other pathogenic parasites tested (Trypanosoma brucei, T. cruzi, Plasmodium falciparum, Pneumocystis carinii, and Toxoplasma gondii).


Subject(s)
DNA, Protozoan/chemistry , Leishmania donovani/genetics , Leishmaniasis, Visceral/parasitology , Polymerase Chain Reaction , Repetitive Sequences, Nucleic Acid , Animals , Base Sequence , Blotting, Southern , Cloning, Molecular , DNA Probes , DNA, Protozoan/isolation & purification , Dogs , Electrophoresis, Polyacrylamide Gel , Genetic Vectors , Humans , Leishmaniasis, Visceral/diagnosis , Molecular Sequence Data , Oligonucleotides/chemistry , Restriction Mapping , Sensitivity and Specificity , Species Specificity
19.
Oncogene ; 8(5): 1195-202, 1993 May.
Article in English | MEDLINE | ID: mdl-8386824

ABSTRACT

By successive screenings of cDNA libraries prepared from human tumours and from human foreskin keratinocytes, we have isolated overlapping cDNAs coding for a novel protein which we call Ron, with sequence characteristics of a receptor protein tyrosine kinase. Ron is a 1400 amino acid protein structurally similar to the 1408 amino acid product of the C-MET proto-oncogene, the receptor for hepatocyte growth factor and scatter factor. The two proteins have 63% overall sequence identity in their intracellular regions. We have localised the RON gene to human chromosome region 3p21, a region frequently deleted in small cell carcinoma of the lung and in renal cell carcinoma, and which is believed to harbour unidentified tumour suppressor genes. Interestingly, normal lung tissue contains transcripts of the RON gene.


Subject(s)
Protein-Tyrosine Kinases/genetics , Proto-Oncogene Proteins/genetics , Receptor Protein-Tyrosine Kinases , Receptors, Cell Surface/genetics , Amino Acid Sequence , Base Sequence , Cell Line , Chromosome Mapping , Chromosomes, Human, Pair 3 , DNA/chemistry , DNA/genetics , DNA/isolation & purification , Gene Expression , Humans , Molecular Sequence Data , Protein-Tyrosine Kinases/chemistry , Proto-Oncogene Mas , Proto-Oncogene Proteins/chemistry , Proto-Oncogene Proteins c-met , Receptors, Cell Surface/chemistry
20.
Genomics ; 16(1): 248-51, 1993 Apr.
Article in English | MEDLINE | ID: mdl-8486366

ABSTRACT

The selective amplification by polymerase chain reaction (PCR) of gene fragments corresponding to new G-protein-coupled receptors resulted in the cloning of 18 orphan members of this gene family. Of these, three human clones amplified from genomic DNA (HGMP03, HGMP04, and HGMP05) were shown to be structurally related. Genomic clones corresponding to HGMP03 and HGMP05 were isolated and their putative coding region sequenced. Following the characterization of two interleukin 8 (IL-8) receptors, HGMP03 appeared to encode the high-affinity IL-8 receptor, whereas the partial clone HGMP04 encodes the low-affinity IL-8 receptor. Comparison with the cDNA sequence suggests that the high-affinity receptor gene is split by an intron in the 5' untranslated region. The high-affinity receptor gene was mapped by in situ hybridization to the 2q33-q36 region of the human genome. The HGMP05 locus turned out to be a pseudogene for the low-affinity IL-8 receptor (87% identity), with multiple frameshifts and point mutations introducing stop codons. Southern blotting on genomic DNA did not allow the further detection of related loci in the human genome.


Subject(s)
Chromosomes, Human, Pair 2 , Pseudogenes , Receptors, Immunologic/genetics , Amino Acid Sequence , Base Sequence , Chromosome Mapping , Cloning, Molecular , DNA/genetics , Genome, Human , Humans , In Situ Hybridization , Molecular Sequence Data , Multigene Family , Polymerase Chain Reaction , Receptors, Immunologic/metabolism , Receptors, Interleukin-8A , Sequence Homology, Nucleic Acid
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