RÉSUMÉ
NADK2 encodes the mitochondrial form of nicotinamide adenine dinucleotide (NAD) kinase, which phosphorylates NAD. Rare recessive mutations in human NADK2 are associated with a syndromic neurological mitochondrial disease that includes metabolic changes, such as hyperlysinemia and 2,4 dienoyl CoA reductase (DECR) deficiency. However, the full pathophysiology resulting from NADK2 deficiency is not known. Here, we describe two chemically induced mouse mutations in Nadk2-S326L and S330P-which cause severe neuromuscular disease and shorten lifespan. The S330P allele was characterized in detail and shown to have marked denervation of neuromuscular junctions by 5 weeks of age and muscle atrophy by 11 weeks of age. Cerebellar Purkinje cells also showed progressive degeneration in this model. Transcriptome profiling on brain and muscle was performed at early and late disease stages. In addition, metabolomic profiling was performed on the brain, muscle, liver and spinal cord at the same ages and on plasma at 5 weeks. Combined transcriptomic and metabolomic analyses identified hyperlysinemia, DECR deficiency and generalized metabolic dysfunction in Nadk2 mutant mice, indicating relevance to the human disease. We compared findings from the Nadk model to equivalent RNA sequencing and metabolomic datasets from a mouse model of infantile neuroaxonal dystrophy, caused by recessive mutations in Pla2g6. This enabled us to identify disrupted biological processes that are common between these mouse models of neurological disease, as well as those processes that are gene-specific. These findings improve our understanding of the pathophysiology of neuromuscular diseases and describe mouse models that will be useful for future preclinical studies.
Sujet(s)
Hyperlysinémies , Dystrophies neuroaxonales , Animaux , Souris , Humains , NAD/génétique , Dystrophies neuroaxonales/génétique , Dystrophies neuroaxonales/métabolisme , Modèles animaux de maladie humaine , Expression des gènes , Phosphotransferases (Alcohol Group Acceptor)/génétique , Protéines mitochondriales/génétique , Group VI Phospholipases A2/génétiqueRÉSUMÉ
Epstein-Barr virus (EBV) is associated with a variety of tumors and nonmalignant conditions. Latent EBV genomes in cells, including tumor cells, are often CpG methylated, whereas virion DNA is not CpG methylated. We demonstrate that methyl CpG binding magnetic beads can be used to fractionate among sources of EBV DNA (DNA extracted from laboratory-purified virions vs DNA extracted from latently infected cell lines). We then applied the technique to plasma specimens and showed that this technique can distinguish EBV DNA from patients with EBV-associated tumors (nasopharyngeal carcinoma, Hodgkin lymphoma) and viral DNA from patients without EBV-associated tumors, including immunocompromised patients and patients with EBV(-) Hodgkin lymphoma.
Sujet(s)
Infections à virus Epstein-Barr , Maladie de Hodgkin , ADN viral/métabolisme , Herpèsvirus humain de type 4/génétique , Herpèsvirus humain de type 4/métabolisme , Humains , MéthylationRÉSUMÉ
The Unique Minds Program (Stern, Unique Minds Program, 1999) addresses the socio-emotional needs of children with learning disabilities (LD) and their families. Children and their parents work together in a multiple family group to learn more about LD and themselves as people with the capacity to solve problems in a collaborative way, including problems in family school relationships. This article reports the cultural adaptation of the program for use in Spain and findings from a feasibility study involving three multiple family groups and a total of 15 children and 15 mothers, using a pre-post design. This Spanish adaptation of the program is called "Mentes Únicas". Standardized outcome measures indicated an overall statistically significant decrease in children's self-rated maladjustment and relationship difficulties by the end of the program. Improvements were endorsed by most mothers, although they were not always recognized by the children's teachers. The program had a high level of acceptability: Mothers and children felt safe, understood, and helped throughout the sessions. The efficacy of the adapted intervention for the context of Spain remains to be tested in a more rigorous study.
Sujet(s)
Thérapie familiale/méthodes , Incapacités d'apprentissage/psychologie , Incapacités d'apprentissage/thérapie , Mères/psychologie , Adulte , Enfant , Compétence culturelle , Émotions , Études de faisabilité , Femelle , Connaissances, attitudes et pratiques en santé , Humains , Mâle , Adulte d'âge moyen , Relations parent-enfant , Acceptation des soins par les patients , Concept du soi , Comportement social , EspagneRÉSUMÉ
Primary open angle glaucoma (POAG) is a genetically and phenotypically complex disease that is a leading cause of blindness worldwide. Previously we completed a genome-wide scan for early-onset POAG that identified a locus on 9q22 (GLC1J). To identify potential causative variants underlying GLC1J, we used targeted DNA capture followed by high throughput sequencing of individuals from four GLC1J pedigrees, followed by Sanger sequencing to screen candidate variants in additional pedigrees. A mutation likely to cause early-onset glaucoma was not identified, however COL15A1 variants were found in the youngest affected members of 7 of 15 pedigrees with variable disease onset. In addition, the most common COL15A1 variant, R163H, influenced the age of onset in adult POAG cases. RNA in situ hybridization of mouse eyes shows that Col15a1 is expressed in the multiple ocular structures including ciliary body, astrocytes of the optic nerve and cells in the ganglion cell layer. Sanger sequencing of COL18A1, a related multiplexin collagen, identified a rare variant, A1381T, in members of three additional pedigrees with early-onset disease. These results suggest genetic variation in COL15A1 and COL18A1 can modify the age of onset of both early and late onset POAG.
Sujet(s)
Collagène de type XVIII/génétique , Collagène/génétique , Variation génétique , Glaucome à angle ouvert/génétique , Adulte , Âge de début , Sujet âgé , Animaux , Exons , Femelle , Génotype , Humains , Mâle , Souris , Adulte d'âge moyen , Pedigree , Polymorphisme de nucléotide simpleRÉSUMÉ
BACKGROUND: Porencephaly (cystic cavities of the brain) is caused by perinatal vascular accidents from various causes. Several familial cases have been described and autosomal dominant inheritance linked to chromosome 13q has been suggested. COL4A1 is an essential component in basal membrane stability. Mouse mutants bearing an in-frame deletion of exon 40 of Col4a1 either die from haemorrhage in the perinatal period or have porencephaly in survivors. A report of inherited mutations in COL4A1 in two families has shown that familial porencephaly may have the same cause in humans. OBJECTIVE: To describe three novel COL4A1 mutations. RESULTS: The three mutations occurred in three unrelated Dutch families. There were two missense mutations of glycine residues predicted to result in abnormal collagen IV assembly, and one mutation predicted to abolish the traditional COL4A1 start codon. The last mutation was also present in an asymptomatic obligate carrier with white matter abnormalities on brain magnetic resonance imaging. CONCLUSIONS: This observation confirms COL4A1 as a major locus for genetic predisposition to perinatal cerebral haemorrhage and porencephaly and suggests variable expression of COL4A1 mutations.
Sujet(s)
Encéphalopathies/génétique , Collagène de type IV/génétique , Adulte , Encéphalopathies/diagnostic , Encéphalopathies/anatomopathologie , Enfant , Enfant d'âge préscolaire , Collagène de type IV/composition chimique , Collagène de type IV/physiologie , Analyse de mutations d'ADN , Femelle , Humains , Nourrisson , Mâle , Adulte d'âge moyen , Mutation faux-sens , Pedigree , Structure tertiaire des protéinesRÉSUMÉ
BACKGROUND: Glaucoma is a blinding disease usually associated with high intraocular pressure (IOP). In some families, abnormal anterior segment development contributes to glaucoma. The genes causing anterior segment dysgenesis and glaucoma in most of these families are not identified and the affected developmental processes are poorly understood. Bone morphogenetic proteins (BMPs) participate in various developmental processes. We tested the importance of Bmp4 gene dosage for ocular development and developmental glaucoma. RESULTS: Bmp4+/- mice have anterior segment abnormalities including malformed, absent or blocked trabecular meshwork and Schlemm's canal drainage structures. Mice with severe drainage structure abnormalities, over 80% or more of their angle's extent, have elevated IOP. The penetrance and severity of abnormalities is strongly influenced by genetic background, being most severe on the C57BL/6J background and absent on some other backgrounds. On the C57BL/6J background there is also persistence of the hyaloid vasculature, diminished numbers of inner retinal cells, and absence of the optic nerve. CONCLUSIONS: We demonstrate that heterozygous deficiency of BMP4 results in anterior segment dysgenesis and elevated IOP. The abnormalities are similar to those in human patients with developmental glaucoma. Thus, BMP4 is a strong candidate to contribute to Axenfeld-Rieger anomaly and other developmental conditions associated with human glaucoma. BMP4 also participates in posterior segment development and wild-type levels are usually critical for optic nerve development on the C57BL/6J background. Bmp4+/- mice are useful for studying various components of ocular development, and may allow identification of strain specific modifiers affecting a variety of ocular phenotypes.
Sujet(s)
Pôle antérieur du bulbe oculaire/croissance et développement , Protéines morphogénétiques osseuses/génétique , Protéines morphogénétiques osseuses/physiologie , Pression intraoculaire , Hypertension oculaire/étiologie , Animaux , Pôle antérieur du bulbe oculaire/malformations , Protéine morphogénétique osseuse de type 4 , Électrorétinographie , Malformations oculaires/étiologie , Malformations oculaires/anatomopathologie , Dosage génique , Hétérozygote , Souris , Souris de lignée C57BL , Hypertension oculaire/anatomopathologie , Nerf optique/croissance et développement , Phénotype , Vaisseaux rétiniens/croissance et développementRÉSUMÉ
Glaucoma is a heterogeneous eye disease and a major cause of blindness worldwide. Recently, primary open angle glaucoma (POAG)-associated mutations have been found in the trabecular meshwork inducible glucocorticoid response gene (TIGR), also known as the myocilin gene (MYOC), at the GLC1A locus on chromosome 1q21-q31. These mutations occurred in a subset of patients with juvenile- and adult-onset POAG and exhibited autosomal dominant inheritance. Ocular expression and its involvement in POAG suggest that TIGR/MYOC may have a role(s) in regulating intraocular pressure (IOP). Here, we report the generation and analysis of mice heterozygous and homozygous for a targeted null mutation in Myoc. Our study shows that Myoc mutant mice are both viable and fertile. Our in vivo findings further demonstrate that Myoc is not required for normal IOP or normal ocular morphology. The lack of a discernable phenotype in both Myoc-heterozygous and Myoc-null mice suggests that haploinsufficiency is not a critical mechanism for POAG in individuals with mutations in MYOC. Instead, disease-causing mutations in humans likely act by gain of function.
Sujet(s)
Protéines de l'oeil/physiologie , Glaucome à angle ouvert/anatomopathologie , Glycoprotéines/physiologie , Animaux , Protéines du cytosquelette , Oeil/métabolisme , Oeil/anatomopathologie , Protéines de l'oeil/génétique , Expression des gènes , Ciblage de gène/méthodes , Glycoprotéines/génétique , Humains , Pression intraoculaire , Souris , Souris de lignée C57BL , Souris knockout , Mutagenèse , ARN messagerRÉSUMÉ
BACKGROUND: Little is known about genetic factors affecting intraocular pressure (IOP) in mice and other mammals. The purpose of this study was to determine the IOPs of genetically distinct mouse strains, assess the effects of factors such as age, sex and time of day on IOP in specific strain backgrounds, and to assess the effects of specific candidate gene mutations on IOP. RESULTS: Based on over 30 studied mouse strains, average IOP ranges from approximately 10 to 20 mmHg. Gender does not typically affect IOP and aging results in an IOP decrease in some strains. Most tested strains exhibit a diurnal rhythm with IOP being the highest during the dark period of the day. Homozygosity for a null allele of the carbonic anhydrase II gene (Car2n) does not alter IOP while homozygosity for a mutation in the leptin receptor gene (Leprdb) that causes obesity and diabetes results in increased IOP. Albino C57BL/6J mice homozygous for a tyrosinase mutation (Tyrc-2J) have higher IOPs than their pigmented counterparts. CONCLUSIONS: Genetically distinct mouse strains housed in the same environment have a broad range of IOPs. These IOP differences are likely due to interstrain genetic differences that create a powerful resource for studying the regulation of IOP. Age, time of day, obesity and diabetes have effects on mouse IOP similar to those in humans and other species. Mutations in two of the assessed candidate genes (Lepr and Tyr) result in increased IOP. These studies demonstrate that mice are a practical and powerful experimental system to study the genetics of IOP regulation and disease processes that raise IOP to harmful levels.
Sujet(s)
Pression intraoculaire , Lignées consanguines de souris , Modèles animaux , Facteurs âges , Anesthésie , Animaux , Pression sanguine , Protéines du cytosquelette , Environnement , Protéines de l'oeil/génétique , Femelle , Variation génétique , Glaucome/génétique , Glycoprotéines/génétique , Pression intraoculaire/génétique , Mâle , Souris , Lignées consanguines de souris/génétique , Lignées consanguines de souris/physiologie , Monophenol monooxygenase/déficit , Mutation , Périodicité , Rats , Reproductibilité des résultats , Facteurs de risque , Facteurs sexuels , Spécificité d'espèce , Facteurs tempsRÉSUMÉ
BACKGROUND: The iridocorneal angle forms in the mammalian eye from undifferentiated mesenchyme between the root of the iris and cornea. A major component is the trabecular meshwork, consisting of extracellular matrix organized into a network of beams, covered in trabecular endothelial cells. Between the beams, channels lead to Schlemm's canal for the drainage of aqueous humor from the eye into the blood stream. Abnormal development of the iridocorneal angle that interferes with ocular fluid drainage can lead to glaucoma in humans. Little is known about the precise mechanisms underlying angle development. There are two main hypotheses. The first proposes that morphogenesis involves mainly cell differentiation, matrix deposition and assembly of the originally continuous mesenchymal mass into beams, channels and Schlemm's canal. The second, based primarily on rat studies, proposes that cell death and macrophages play an important role in forming channels and beams. Mice provide a potentially useful model to understand the origin and development of angle structures and how defective development leads to glaucoma. Few studies have assessed the normal structure and development of the mouse angle. We used light and electron microscopy and a cell death assay to define the sequence of events underlying formation of the angle structures in mice. RESULTS: The mouse angle structures and developmental sequence are similar to those in humans. Cell death was not detectable during the period of trabecular channel and beam formation. CONCLUSIONS: These results support morphogenic mechanisms involving organization of cellular and extracellular matrix components without cell death or atrophy.
Sujet(s)
Chambre antérieure du bulbe oculaire/cytologie , Chambre antérieure du bulbe oculaire/embryologie , Réseau trabéculaire de la sclère/cytologie , Réseau trabéculaire de la sclère/embryologie , Animaux , Chambre antérieure du bulbe oculaire/croissance et développement , Chambre antérieure du bulbe oculaire/ultrastructure , Mort cellulaire/physiologie , Cornée/cytologie , Cornée/embryologie , Cornée/croissance et développement , Cornée/ultrastructure , Matrice extracellulaire/physiologie , Matrice extracellulaire/ultrastructure , Humains , Iris/cytologie , Iris/embryologie , Iris/croissance et développement , Iris/ultrastructure , Souris , Souris de lignée A , Souris de lignée BALB C , Souris de lignée C3H , Souris de lignée C57BL , Souris de lignée DBA , Souris de lignée MRL lpr , Lignées consanguines de souris , Microscopie électronique à balayage/méthodes , Réseau trabéculaire de la sclère/croissance et développement , Réseau trabéculaire de la sclère/ultrastructureRÉSUMÉ
Hyperhomocysteinemia, a risk factor for cardiovascular disease, is caused by nutritional and/or genetic disruptions in homocysteine metabolism. The most common genetic cause of hyperhomocysteinemia is the 677C-->T mutation in the methylenetetrahydrofolate reductase (MTHFR) gene. This variant, with mild enzymatic deficiency, is associated with an increased risk for neural tube defects and pregnancy complications and with a decreased risk for colon cancer and leukemia. Although many studies have reported that this variant is also a risk factor for vascular disease, this area of investigation is still controversial. Severe MTHFR deficiency results in homocystinuria, an inborn error of metabolism with neurological and vascular complications. To investigate the in vivo pathogenetic mechanisms of MTHFR deficiency, we generated mice with a knockout of MTHFR: Plasma total homocysteine levels in heterozygous and homozygous knockout mice are 1.6- and 10-fold higher than those in wild-type littermates, respectively. Both heterozygous and homozygous knockouts have either significantly decreased S-adenosylmethionine levels or significantly increased S-adenosylhomocysteine levels, or both, with global DNA hypomethylation. The heterozygous knockout mice appear normal, whereas the homozygotes are smaller and show developmental retardation with cerebellar pathology. Abnormal lipid deposition in the proximal portion of the aorta was observed in older heterozygotes and homozygotes, alluding to an atherogenic effect of hyperhomocysteinemia in these mice.
Sujet(s)
Aorte/métabolisme , Hyperhomocystéinémie/génétique , Métabolisme lipidique , Système nerveux/anatomopathologie , Oxidoreductases acting on CH-NH group donors/physiologie , Animaux , Séquence nucléotidique , Méthylation de l'ADN , Amorces ADN , Hétérozygote , Homozygote , Hyperhomocystéinémie/enzymologie , Hyperhomocystéinémie/anatomopathologie , Methylenetetrahydrofolate reductase (NADPH2) , Souris , Souris knockout , Oxidoreductases acting on CH-NH group donors/génétique , Oxidoreductases acting on CH-NH group donors/métabolisme , Phénotype , RT-PCRRÉSUMÉ
BACKGROUND: Glaucoma is a common disease but its molecular etiology is poorly understood. It involves retinal ganglion cell death and optic nerve damage that is often associated with elevated intraocular pressure. Identifying genes that modify glaucoma associated phenotypes is likely to provide insights to mechanisms of glaucoma. We previously reported glaucoma in DBA/2J mice caused by recessive alleles at two loci, isa and ipd, that cause iris stromal atrophy and iris pigment dispersion, respectively. A approach for identifying modifier genes is to study the effects of specific mutations in different mouse strains. When the phenotypic effect of a mutation is modified upon its introduction into a new strain, crosses between the parental strains can be used to identify modifier genes. The purpose of this study was to determine if the effects of the DBA/2J derived isa and ipd loci are modified in strain AKXD-28/Ty. RESULTS: AKXD-28/Ty mice develop glaucoma characterized by intraocular pressure elevation, retinal ganglion loss, and optic nerve excavation. In AKXD-28/Ty, isa causes an iris stromal atrophy phenotype as in DBA/2J. However, the iris pigment dispersion phenotype associated with ipd in DBA/2J does not occur in AKXD-28/Ty. Additionally, a greater severity and speed of retinal and optic nerve damage following intraocular pressure elevation in AKXD-28/Ty compared to DBA/2J mice suggests that AKXD-28/Ty is more susceptible to pressure-induced cell death. CONCLUSIONS: The consequences of the ipd and isa mutations are modified in the AKXD-28/Ty background. These strains provide a resource for the identification of modifier genes that modulate pigment dispersion and susceptibility to pressure-induced cell death.
Sujet(s)
Prédisposition génétique à une maladie , Glaucome/génétique , Glaucome/anatomopathologie , Animaux , Atrophie , Femelle , Glaucome/diagnostic , Iris/anatomopathologie , Mâle , Souris , Souris de lignée C57BL , Souris de lignée DBA , Lignées consanguines de souris , Mutation , Atteintes du nerf optique/génétique , Atteintes du nerf optique/anatomopathologie , Phénotype , Épithélium pigmentaire de l'oeil/anatomopathologie , Rétinopathies/génétique , Rétinopathies/anatomopathologie , Facteurs sexuels , Spécificité d'espèceRÉSUMÉ
BACKGROUND: The atrial natriuretic peptide (ANP) family is a complex system consisting of at least three polypeptides and at least three types of receptor. Each peptide interacts with different types of receptor at varying degrees of affinity. To determine if natriuretic peptide levels influence natriuretic peptide receptor expression and regulation, we examined the expression of guanylyl cyclase linked GC-A, GC-B and C-receptor in the lungs of mice with a mutation that inactivates the ANP gene (Nppa). METHODS: The mRNA level of GC-A, GC-B and C-receptor in the lung were studied by ribonuclease protection assays (RPA). RESULTS: Results of RPA showed that although the mRNA level of GC-A and GC-B of heterozygous ANP+/- was not different from wild type ANP+/+ mice, they were significantly higher in the homozygous mutant ANP-/- mice. In addition, C-receptor mRNA level in ANP+/- and ANP-/- was significantly lower than ANP+/+ mice. The C-receptor results were confirmed by receptor binding assays and affinity cross-linking studies. CONCLUSIONS: Taken together these data suggest that permanent removal of ANP from the natriuretic peptide system results in an up-regulation of GC-A and GC-B, and a corresponding down-regulation of C-receptor in the lung of proANP gene disrupted mice. We postulated that changes in the natriuretic peptide receptor population may result in chronic hypertension and cardiac hypertrophy in the ANP-/- mice.
Sujet(s)
Facteur atrial natriurétique/génétique , Hypertension artérielle/métabolisme , Poumon/métabolisme , Récepteur facteur natriurétique auriculaire/métabolisme , Analyse de variance , Animaux , Autoradiographie , Expression des gènes , Guanylate cyclase/génétique , Souris , Souris de lignée C57BL , Souches mutantes de souris , ARN messager/analyse , Récepteur facteur natriurétique auriculaire/génétiqueRÉSUMÉ
Anterior segment developmental disorders, including Axenfeld-Rieger anomaly (ARA), variably associate with harmfully elevated intraocular pressure (IOP), which causes glaucoma. Clinically observed dysgenesis does not correlate with IOP, however, and the etiology of glaucoma development is not understood. The forkhead transcription factor genes Foxc1 (formerly Mf1 ) and Foxc2 (formerly Mfh1 ) are expressed in the mesenchyme from which the ocular drainage structures derive. Mutations in the human homolog of Foxc1, FKHL7, cause dominant anterior segment defects and glaucoma in various families. We show that Foxc1 (+/-)mice have anterior segment abnormalities similar to those reported in human patients. These abnormalities include small or absent Schlemm's canal, aberrantly developed trabecular meshwork, iris hypoplasia, severely eccentric pupils and displaced Schwalbe's line. The penetrance of clinically obvious abnormalities varies with genetic background. In some affected eyes, collagen bundles were half normal diameter, or collagen and elastic tissue were very sparse. Thus, abnormalities in extracellular matrix synthesis or organization may contribute to development of the ocular phenotypes. Despite the abnormalities in ocular drainage structures in Foxc1 (+/-)mice, IOP was normal in almost all mice analyzed, on all genetic backgrounds and at all ages. Similar abnormalities were found in Foxc2 (+/-)mice, but no disease-associated mutations were identified in the human homolog FKHL14 in 32 ARA patients. Foxc1 (+/-)and Foxc2 (+/-)mice are useful models for studying anterior segment development and its anomalies, and may allow identification of genes that interact with Foxc1 and Foxc2 (or FKHL7 and FKHL14 ) to produce a phenotype with elevated IOP and glaucoma.
Sujet(s)
Pôle antérieur du bulbe oculaire/malformations , Protéines de liaison à l'ADN/génétique , Oeil/embryologie , Facteurs de transcription/génétique , Animaux , Corps ciliaire/malformations , Protéines de liaison à l'ADN/physiologie , Facteurs de transcription Forkhead , Génotype , Glaucome/génétique , Haplotypes , Hétérozygote , Humains , Hybridation in situ , Pression intraoculaire/génétique , Souris , Souches mutantes de souris , Microscopie électronique , Mutagenèse , Phénotype , Spécificité d'espèce , Facteurs de transcription/physiologieRÉSUMÉ
The drive to characterize functions of human genes on a global scale has stimulated interest in large-scale generation of mouse mutants. Conventional germ-cell mutagenesis with N-ethyl-N-nitrosourea (ENU) is compromised by an inability to monitor mutation efficiency, strain and interlocus variation in mutation induction, and extensive husbandry requirements. To overcome these obstacles and develop new methods for generating mouse mutants, we devised protocols to generate germline chimaeric mice from embryonic stem (ES) cells heavily mutagenized with ethylmethanesulphonate (EMS). Germline chimaeras were derived from cultures that underwent a mutation rate of up to 1 in 1,200 at the Hprt locus (encoding hypoxanthine guanine phosphoribosyl transferase). The spectrum of mutations induced by EMS and the frameshift mutagen ICR191 was consistent with that observed in other mammalian cells. Chimaeras derived from ES cells treated with EMS transmitted mutations affecting several processes, including limb development, hair growth, hearing and gametogenesis. This technology affords several advantages over traditional mutagenesis, including the ability to conduct shortened breeding schemes and to screen for mutant phenotypes directly in ES cells or their differentiated derivatives.
Sujet(s)
Malformations dues aux médicaments et aux drogues/génétique , Malformations multiples/génétique , Méthanesulfonate d'éthyle/toxicité , 1-Éthyl-1-nitroso-urée/toxicité , Souches mutantes de souris/génétique , Mutagenèse , Mutagènes/toxicité , Cellules souches/effets des médicaments et des substances chimiques , Malformations multiples/induit chimiquement , Animaux , Os et tissu osseux/malformations , Chimère/génétique , Femelle , Gènes létaux , Hypoxanthine phosphoribosyltransferase/génétique , Anomalies morphologiques congénitales des membres/génétique , Mâle , Souris , Souris de lignée C57BL , Mutation ponctuelle , Épissage des ARN , Rétine/malformations , Testicule/malformationsRÉSUMÉ
Ocular neovascularization is the leading cause of blindness in developed countries and often causes rapid loss of vision in age-related macular degeneration. Acute visual loss is most often due to hemorrhage from new vessels that have extended from the choroid into the subretinal space. Growth of abnormal vessels beneath the retina in this condition is known as subretinal neovascularization (SRN). Age-related animal models of macular degeneration and SRN have not been described. Current animal models of SRN depend on chemical or physical stimuli to initiate growth of subretinal vessels. The genes responsible for age-related human macular degeneration with SRN have not been firmly identified. We report an angiogenic phenotype in Bst/+ mice that is age-related, clinically evident, and resembles human SRN. This represents a spontaneous, genetically determined model of SRN. Bst/+ mice offer the possibility of exploring the molecular mechanisms of SRN without the need for exogenous agents.
Sujet(s)
Vieillissement/physiologie , Néovascularisation rétinienne/génétique , Animaux , Cartographie chromosomique , Modèles animaux de maladie humaine , Oeil/anatomopathologie , Angiographie fluorescéinique , Souris , Souris de lignée C57BL , Néovascularisation rétinienne/anatomopathologieRÉSUMÉ
Gene characterization holds great promise for understanding molecular mechanisms of disease. Although glaucoma gene identification is very valuable and allows assessment of an individual's genetic risk, it is not by itself sufficient to answer detailed questions about pathogenesis. Despite the recent identification of a number of glaucoma genes, there are still many questions regarding the ways in which mutations in these genes cause disease. The mouse system, including the ability to alter specific genes, provides a powerful experimental system for hypothesis testing and molecular dissection of pathogenesis subsequent to gene identification. The ability to control both genetic and environmental factors will allow the use of mice to identify modifier genes that alter complex glaucoma phenotypes and that are especially difficult to identify in human families. By providing a bridge between gene identification and tests of gene function, mouse studies will be an important complement to those in humans and other species. This article summarizes the recent use of mice and the future potential of applying approaches of mouse genetics to intraocular pressure and glaucoma research.
Sujet(s)
Modèles animaux de maladie humaine , Glaucome/génétique , Souris , Animaux , Cartographie chromosomique , Glaucome/anatomopathologie , Humains , Pression intraoculaire/génétiqueRÉSUMÉ
PURPOSE: Mice are an increasingly important tool in ophthalmic research. As a result of studying spontaneous and induced mutations, many new ocular diseases have been described in mice in recent years, including several degenerative retinal diseases that demonstrate progression with age. Clearly, documentation of progressive changes in clinical phenotype is an important facet of characterizing new mutations and for comparing them with human diseases. Despite these facts, there are few published photographs of mouse fundi. The small size of the mouse eye and the steep curvature of its structures have made it difficult to obtain high quality fundus photographs. The purpose of this work was to develop procedures for mouse fundus photography and angiography and to use these techniques to examine several new mouse strains with ocular abnormalities. METHODS: We have used a small animal fundus camera and condensing lens to develop a reliable technique for producing high quality fundus images of conscious albino and pigmented mice. The fundus camera also was utilized to develop a method for fluorescein angiography, which demonstrated the normal retinal vascular bed as well as abnormal vascular leakage. In addition, several mouse strains with previously unreported ocular abnormalities (including two with inherited optic nerve colobomas) and a catalogue of previously unpublished clinical images for various mutant mice are presented. RESULTS: Altogether, we provide clinical images for C57BL/6J, BALB/cByJ, retinal degeneration 1 (rd1), Rd2, rd3, rd7, achondroplasia, nervous, motor neuron degeneration, Purkinje cell degeneration, kidney and retinal defects, optic nerve coloboma 1, and two apparently multigenic optic nerve colobomas in a strain of mixed derivation (ONC) and the inbred CALB/Rk strain. CONCLUSIONS: Our photography procedure reliably produces high quality images of the mouse fundus. This permitted us to record progressive retinal changes over time in the same animal, allowed us to compare the phenotypes of newly discovered retinal mutants to existing mutants at other institutions and to potentially similar human conditions, and finally, permitted us to produce a catalogue of previously unpublished clinical phenotypes for various mutant mice.
Sujet(s)
Angiographie fluorescéinique/méthodes , Fond de l'oeil , Ophtalmoscopes , Photographie (méthode)/méthodes , Vaisseaux rétiniens/anatomie et histologie , Animaux , Souris , Souris de lignée BALB C , Souris de lignée C57BL , Lignées consanguines de sourisRÉSUMÉ
Glaucomas are a major cause of blindness. Visual loss typically involves retinal ganglion cell death and optic nerve atrophy subsequent to a pathologic elevation of intraocular pressure (IOP). Some human glaucomas are associated with anterior segment abnormalities such as pigment dispersion syndrome (PDS) and iris atrophy with associated synechiae. The primary causes of these abnormalities are unknown, and their aetiology is poorly understood. We recently characterized a mouse strain (DBA/2J) that develops glaucoma subsequent to anterior segment changes including pigment dispersion and iris atrophy. Using crosses between mouse strains DBA/2J (D2) and C57BL/6J (B6), we now show there are two chromosomal regions that contribute to the anterior segment changes and glaucoma. Progeny homozygous for the D2 allele of one locus on chromosome 6 (called ipd) develop an iris pigment dispersion phenotype similar to human PDS. ipd resides on a region of mouse chromosome 6 with conserved synteny to a region of human chromosome 7q that is associated with human PDS. Progeny homozygous for the D2 allele of a different locus on chromosome 4 (called isa) develop an iris stromal atrophy phenotype (ISA). The Tyrpl gene is a candidate for isa and likely causes ISA via a mechanism involving pigment production. Progeny homozygous for the D2 alleles of both ipd and isa develop an earlier onset and more severe disease involving pigment dispersion and iris stromal atrophy.
Sujet(s)
Glaucome/génétique , Maladies de l'iris/génétique , Iris/anatomopathologie , Glycoprotéines membranaires , Souris de lignée DBA/génétique , Oxidoreductases , Facteurs âges , Animaux , Atrophie , Cartographie chromosomique , Croisements génétiques , Homozygote , Maladies de l'iris/anatomopathologie , Souris , Souris de lignée BALB C , Souris de lignée C57BL , Lignées consanguines de souris , Répétitions microsatellites , Épithélium pigmentaire de l'oeil/anatomopathologie , Protéines/génétique , Spécificité d'espèceRÉSUMÉ
PURPOSE: To characterize ocular abnormalities associated with iris atrophy in DBA/2J mice and to determine whether mice of this strain develop elevated intraocular pressure (IOP) and glaucoma. METHODS: Different approaches, including slit-lamp biomicroscopy, ophthalmoscopic examination, ultrasound backscatter microscopy, and histology were used to examine the eyes of DBA/2J mice ranging from 2 to 30 months old. IOP was measured in DBA/2J mice of different ages. RESULTS: DBA/2J mice were found to develop pigment dispersion, iris transillumination, iris atrophy, anterior synechias, and elevated IOP. IOP was elevated in most mice by the age of 9 months. These changes were followed by the death of retinal ganglion cells, optic nerve atrophy, and optic nerve cupping. The prevalence and severity of these lesions increased with age. Optic nerve atrophy and optic nerve cupping was present in the majority of mice by the age of 22 months. CONCLUSIONS: DBA/2J mice develop a progressive form of secondary angle-closure glaucoma that appears to be initiated by iris atrophy and the associated formation of synechias. This mouse strain represents a useful model to evaluate mechanisms of pressure-related ganglion cell death and optic nerve atrophy, and to evaluate strategies for neuroprotection.
Sujet(s)
Glaucome capsulaire/anatomopathologie , Maladies héréditaires de l'oeil/anatomopathologie , Glaucome à angle fermé/anatomopathologie , Iris/anatomopathologie , Vieillissement/anatomopathologie , Animaux , Pôle antérieur du bulbe oculaire/anatomopathologie , Atrophie , Mort cellulaire , Modèles animaux de maladie humaine , Évolution de la maladie , Glaucome capsulaire/étiologie , Glaucome capsulaire/génétique , Maladies héréditaires de l'oeil/étiologie , Maladies héréditaires de l'oeil/génétique , Femelle , Glaucome à angle fermé/étiologie , Glaucome à angle fermé/génétique , Pression intraoculaire , Mâle , Souris , Souris de lignée DBA , Hypertension oculaire/étiologie , Hypertension oculaire/génétique , Hypertension oculaire/anatomopathologie , Atrophie optique/étiologie , Atrophie optique/anatomopathologie , Cellules ganglionnaires rétiniennes/anatomopathologieRÉSUMÉ
Activation of the natriuretic peptide system lowers blood pressure and causes the excretion of salt. Atrial natriuretic peptide and B-type natriuretic peptide are the humoral mediators of this effect; they act primarily by binding to membrane-bound natriuretic peptide receptor A (NPRA) and stimulating its intrinsic guanylate cyclase activity. To study whether genetically determined differences in NPRA expression affect blood pressure we have generated mice with one, two, three, or four copies of the gene encoding NPRA (Npr1 in the mouse). Atrial natriuretic peptide-dependent guanylate cyclase activity ranged progressively from approximately one-half normal in one-copy animals to twice normal in four-copy animals (P < 0.001). On different diets (0.05%, 2%, and 8% NaCl), the blood pressures of F1 male mice having only one copy of Npr1 averaged 9.1 mmHg (1 mmHg = 133 Pa) above those of wild-type two-copy males (P < 0.001), whereas males with three copies of the gene had blood pressures averaging 5.2 mmHg below normal (P < 0.01). The blood pressures of the one-copy F1 animals were significantly higher (by 6.2 mmHg; P < 0.01) on the high-salt than on the low-salt diet. The blood pressures of four-copy F3 males were significantly lower (by 7 mmHg; P < 0.05) on the high-salt than on the low-salt diet. These results demonstrate that below normal Npr1 expression leads to a salt-sensitive increase in blood pressure, whereas above normal Npr1 expression lowers blood pressures and protects against high dietary salt.