RESUMEN
BACKGROUND: Ectopic ureters (EUs) associated with varying combinations of urinary incontinence, hydronephrosis, and urinary tract infection have been identified in related North American Entlebucher Mountain Dogs. OBJECTIVES: To characterize the disease phenotype in affected dogs and evaluate possible modes of inheritance. ANIMALS: Twenty client-owned Entlebucher Mountain Dogs. Nine dogs had clinical signs of urinary tract disease. METHODS: Prospective case series in which 17 dogs were evaluated with excretory urography, ultrasonography, and urethrocystoscopy. Three additional dogs were evaluated by necropsy alone. Clinical and pedigree histories from 165 North American Entlebuchers were compiled for analysis. RESULTS: Eleven female and 2 male dogs were found to have EUs. Six females and 1 male were continent. Bilateral intravesicular ectopic ureters (IVEUs) were identified in 9 dogs, bilateral extravesicular ectopic ureters (EVEUs) in 3 dogs, and 1 dog had IVEU and EVEU. Hydronephrosis was identified in 5 dogs, 3 of which had bilateral IVEUs. Two necropsied dogs had bilateral hydronephrosis with presumed ureterovesical junction obstruction associated with chronic granulation tissue or lymphoplasmacytic inflammation. Twenty-six dogs with EUs were identified in the pedigree. Because of incomplete penetrance, mode of inheritance could not be determined. CONCLUSIONS AND CLINICAL IMPORTANCE: Ureteral ectopia is common in North American Entlebucher Mountain Dogs and clinical signs alone could not reliably predict disease phenotype. EVEUs were associated with urinary incontinence and occasionally hydronephrosis. IVEUs were clinically silent or associated with hydronephrosis. Further analyses are necessary to confirm and characterize the hereditary nature of the disorder.
Asunto(s)
Enfermedades de los Perros/congénito , Enfermedades Ureterales/veterinaria , Incontinencia Urinaria/veterinaria , Animales , Enfermedades de los Perros/genética , Enfermedades de los Perros/patología , Perros , Femenino , Predisposición Genética a la Enfermedad , Masculino , Linaje , Enfermedades Ureterales/congénito , Enfermedades Ureterales/patología , Incontinencia Urinaria/genética , Incontinencia Urinaria/patologíaRESUMEN
BACKGROUND: Congenital, nonepidermolytic cornification disorders phenotypically resembling human autosomal recessive ichthyosis have been described in purebred dog breeds, including Jack Russell terrier (JRT) dogs. One cause of gene mutation important to humans and dogs is transposon insertions. OBJECTIVES: To describe an autosomal recessive, severe nonepidermolytic ichthyosis resembling lamellar ichthyosis (LI) in JRT dogs due to insertion of a long interspersed nucleotide element (LINE-1) in the transglutaminase 1 (TGM1) gene. METHODS: Dogs were evaluated clinically, and skin samples were examined by light and electron microscopy. Phenotypic information and genotyping with a canine microsatellite marker suggested TGM1 to be a candidate gene. Genomic DNA samples and cDNA generated from epidermal RNA were examined. Consequences of the mutation were evaluated by Western blotting, quantitative reverse transcription-polymerase chain reaction (RT-PCR) and enzyme activity from cultured keratinocytes. RESULTS: Affected dogs had generalized severe hyperkeratosis. Histological examination defined laminated to compact hyperkeratosis without epidermolysis; ultrastructurally, cornified envelopes were thin. Affected dogs were homozygous for a 1980-bp insertion within intron 9 of TGM1. The sequence of the insertion was that of a canine LINE-1 element. Quantitative RT-PCR indicated a significant decrease in TGM1 mRNA in affected dogs compared with wild-type. TGM1 protein was markedly decreased on immunoblotting, and membrane-associated enzyme activity was diminished in affected dogs. CONCLUSIONS: Based on morphological and molecular features, this disease is homologous with TGM1-deficient LI in humans, clinically models LI better than the genetically modified mouse and represents its first spontaneous animal model. This is the first reported form of LI due to transposon insertion.
Asunto(s)
Enfermedades de los Perros/genética , Ictiosis Lamelar/veterinaria , Elementos de Nucleótido Esparcido Largo/genética , Mutagénesis Insercional/genética , Transglutaminasas/genética , Animales , Biopsia/veterinaria , Elementos Transponibles de ADN/genética , Enfermedades de los Perros/patología , Perros , Femenino , Marcadores Genéticos , Ictiosis Lamelar/genética , Ictiosis Lamelar/patología , Inmunohistoquímica , Intrones/genética , Masculino , ARN Mensajero/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/veterinaria , Piel/patología , Especificidad de la Especie , Transglutaminasas/deficiencia , Transglutaminasas/metabolismoRESUMEN
REASONS FOR PERFORMING STUDY: Airway inflammation in recurrent airway obstruction (RAO) is triggered by housing affected horses in stables.It has been suggested that RAO is an allergic condition, but innate immune mechanisms are also involved. Fungal products activate innate immune mechanisms through toll-like receptor 2 (TLR2). In human airway epithelium, TLR2 activation leads to interleukin (IL)-8 production. This pathway is negatively regulated by the zinc finger protein A20. This study was performed to enhance understanding of innate immune mechanisms in RAO. HYPOTHESIS: TLR2 and IL-8 mRNA are elevated in RAO during stabling compared with controls. A20 mRNA is negatively associated with the numbers of airway inflammatory cells. OBJECTIVES: To determine TLR2, IL-8 and A20 mRNA expression in lungs of stabled and pastured RAO-affected and control horses. METHODS: Airway obstruction and inflammatory cell counts in bronchoalveolar lavage were measured, and TLR2, IL-8 and A20 mRNA expression quantified by qRT-PCR in 6 RAO-affected and 6 control horses, during and after exposure to hay and straw. RESULTS: Airway obstruction and neutrophils were increased in RAO-affected horses during stabling. While stabling increased IL-8, TLR2 and A20 mRNA were unaffected. TLR2 and A20 were significantly correlated (r = 0.83) and A20 mRNA was negatively associated with inflammatory cells. POTENTIAL RELEVANCE: Stabling does not lead to an increase in TLR2 expression. Other molecules or processes in the TLR2 cascade might be important in fungal-induced airway inflammation. Equine epithelial-derived A20 may be involved in modulation of airway inflammation.
Asunto(s)
Obstrucción de las Vías Aéreas/veterinaria , Bronquios/metabolismo , Enfermedades de los Caballos/fisiopatología , Vivienda para Animales , Receptor Toll-Like 2/genética , Obstrucción de las Vías Aéreas/inmunología , Obstrucción de las Vías Aéreas/fisiopatología , Animales , Bronquios/citología , Bronquios/inmunología , Estudios de Casos y Controles , Femenino , Regulación de la Expresión Génica/fisiología , Enfermedades de los Caballos/inmunología , Caballos , Inmunidad Innata , Inflamación/etiología , Inflamación/inmunología , Inflamación/veterinaria , Interleucina-8/biosíntesis , Interleucina-8/genética , Masculino , Proteínas Nucleares/genética , Proteínas Nucleares/metabolismo , ARN Mensajero/metabolismo , Receptor Toll-Like 2/metabolismoRESUMEN
The upstream promoter sequences of the human carbonic anhydrase II (CA II) gene have been studied by 5' deletion analysis. Promoter activity was assayed by transfection and chloramphenicol acetyltransferase assay in both human HeLa cells and murine L cells. This investigation showed that the CA II promoter is comparable in activity to that of the simian virus 40 early-region promoter and enhancer and that the CA II upstream sequences exert a different pattern of control in the two cell lines.
Asunto(s)
Anhidrasas Carbónicas/genética , Regiones Promotoras Genéticas , Secuencias Reguladoras de Ácidos Nucleicos , Transcripción Genética , Animales , Composición de Base , Codón , ADN Recombinante , Células HeLa , Células L , Ratones , Datos de Secuencia Molecular , Mutación , Plásmidos , Virus 40 de los Simios/genética , TransfecciónRESUMEN
The nucleotide sequence of the 5' region of the human carbonic anhydrase II gene has been determined. This sequence begins 643 base pairs upstream from the ATG start site and continues through exon 1, intron 1, exon 2 and the adjoining 125 nucleotides of intron 2. The human sequence is compared with homologous regions of the mouse (YBR strain) carbonic anhydrase II gene by aligning the two sequences for optimal homology. In addition to a TATA box and a putative CCAAT box (CCACC in human and CCACT in mouse), three conserved tandem-repeat elements in mouse and two in human (consensus: cCNGTCACCTCCgC) are located 15 and 22 base pairs upstream, respectively, from the CCAAT boxes in the human and mouse sequences. This repeat element is similar to a tandem repeat sequence located at about the same position in mammalian beta-globin genes, and may represent regulatory elements common to both the carbonic anhydrase and beta-globin genes. The regions surrounding exon 1 are extremely G + C-rich in both human and mouse genes. In addition, several CCGCCC or GGGCGG sequences which may be important for transcriptional efficiency are found in the 5' flanking regions of the human and mouse genes.
Asunto(s)
Anhidrasas Carbónicas/genética , Genes Reguladores , Genes , Secuencia de Aminoácidos , Animales , Composición de Base , Secuencia de Bases , Clonación Molecular , Enzimas de Restricción del ADN , Cabras , Humanos , Ratones , Conejos , Ratas , Secuencias Repetitivas de Ácidos Nucleicos , Especificidad de la EspecieRESUMEN
A recently reported mRNA, encoding 'carbonic anhydrase-related polypeptide' (CARP) from the Purkinje cells of mouse cerebellum, was shown to have a 30-40% deduced amino acid sequence identity with the carbonic anhydrases (CA) of mammals. In order to compare the mouse and human CARP sequences, we used the polymerase chain reaction (PCR) to amplify human CARP sequences from several cDNA libraries (salivary gland, testis and placenta). The sequence has an 89.3% sequence identity with mouse CARP at the nucleotide level and 97.9% at the amino acid level. This extremely high evolutionary conservation suggests an important function for the CARP gene product.
Asunto(s)
Anhidrasas Carbónicas , Proteínas del Tejido Nervioso/genética , Secuencia de Aminoácidos , Animales , Secuencia de Bases , ADN , Humanos , Ratones , Datos de Secuencia Molecular , Homología de Secuencia de Aminoácido , Homología de Secuencia de Ácido NucleicoRESUMEN
The structure of the gene encoding carbonic anhydrase I (CA I) was determined for the pigtail macaque Macaca nemestrina. When the deduced amino-acid sequence was compared with those of five other primates, four non-primate mammals and a turtle, seven residues were found to be unique and invariant to all of the CA I sequences. A scheme is presented for the probable evolutionary order of the six polymorphic nucleotide changes found in the coding regions of the CA I locus of pigtail macaques.
Asunto(s)
Anhidrasas Carbónicas/genética , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Evolución Biológica , ADN , Humanos , Macaca nemestrina , Ratones , Datos de Secuencia Molecular , Homología de Secuencia de Ácido Nucleico , TortugasRESUMEN
The nucleotide sequence of a clone containing mouse carbonic anhydrase (CA) cDNA in pBR322 has been determined. The cloned cDNA contains all of the coding region except for nucleotides specifying the first eight amino acids, and all of the 3' noncoding region, which consists of 700 nucleotides. A cDNA clone was identified which contains an additional 54 bp at the 5' end, so that the complete amino acid sequence of mouse CA could be deduced. This sequence showed a 73-81% homology with other mammalian CA form II isozymes, 56-63% with form I isozymes, and 52-56% with form III isozymes. By examination of the amino acids which are unique and invariant for each isozyme, the mouse amino acid sequence was found to contain 16 of the 23 residues that are unique and invariant to mammalian CA form II isozymes, but only one or no residue for forms I and III, respectively.
Asunto(s)
Anhidrasas Carbónicas/genética , ADN , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Evolución Biológica , Anhidrasas Carbónicas/biosíntesis , Clonación Molecular , Isoenzimas/genética , Ratones , PlásmidosRESUMEN
The genes encoding carbonic anhydrase I (CA I) have been characterized for chimpanzee (Pan troglodytes) and gorilla (Gorilla gorilla). In addition, 44 nucleotides (nt) at the 5' end of the noncoding first exon (exon 1a), which is unique to the erythroid CA I mRNA, together with 188 nt of the adjacent 5' flanking regions, were sequenced for the corresponding positions of the CA I of orangutan, pigtail macaque, and squirrel monkey. When these 5' flanking regions are compared, along with those published for human and mouse CA I, they were found to contain several conserved sequences that may bind factors involved in the erythroid-specific expression of CA I. Comparisons of the human, chimpanzee, and gorilla coding and noncoding CA I sequences do not significantly deviate from a pattern of trichotomy for the evolutionary origins of these three hominoid species.
Asunto(s)
Anhidrasas Carbónicas/genética , Eritrocitos , Gorilla gorilla/genética , Pan troglodytes/genética , Regiones Promotoras Genéticas , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Evolución Biológica , ADN , Exones , Humanos , Macaca nemestrina , Datos de Secuencia Molecular , Pongo pygmaeus , Saimiri , Homología de Secuencia de Ácido NucleicoRESUMEN
Carbonic anhydrase II (CA II) generates the H+ required for osteoclast-mediated bone resorption in humans. We have developed the human promyelocytic cell line HL-60 as a model system with which to study the osteoclast-specific expression of the CA II gene. Treatment of the cell line with 1,25-dihydroxyvitamin D3 resulted in a dramatic de novo induction of CA II at both the protein and mRNA levels. CA II mRNA was also induced to a lesser extent by 12-O-tetradecanoyl phorbol 13-acetate. Treatment with dimethyl sulfoxide did not increase CA II mRNA. These findings indicate that the HL-60 cell line will be a useful model system to study the osteoclast-specific expression of the CA II gene.
Asunto(s)
Calcitriol/farmacología , Anhidrasas Carbónicas/biosíntesis , Regulación de la Expresión Génica , Osteoclastos , Northern Blotting , Western Blotting , Línea Celular , Electroforesis en Acetato de Celulosa , Inducción Enzimática , Humanos , ARN Mensajero/metabolismo , Acetato de Tetradecanoilforbol/farmacologíaRESUMEN
Genes located on human chromosome 12 (HSA12) are conserved on pig chromosomes 5 and 14 (SSC5 and SSC14), with HSA12q23.3-->q24.11 harboring the evolutionary breakpoint between these chromosomes. For this study, pig sequence-tagged sites (STS) were developed for nine HSA12 genes flanking this breakpoint. Radiation hybrid (RH) mapping using the IMpRH panel revealed that COL2A1, DUSP6, KITLG, PAH and STAB2 map to SSC5, while PXN, PLA2G1B, SART3 and TCF1 map to SSC14. Polymorphisms identified in COL2A1, DUSP6, PAH, PLA2G1B and TCF1 were used for genetic linkage mapping and confirmed the map locations for these genes. Our results indicate that the HSA12 evolutionary breakpoint occurs between STAB2 and SART3 in a region spanning less than five million basepairs. These results refine the comparative map of the HSA12 evolutionary breakpoint region and help to further elucidate the extensive gene order rearrangements between HSA12 and SSC5 and 14.
Asunto(s)
Rotura Cromosómica/genética , Mapeo Cromosómico/métodos , Mapeo Cromosómico/veterinaria , Cromosomas Humanos Par 12/genética , Cromosomas/genética , Evolución Molecular , Genes/genética , Porcinos/genética , Animales , Análisis Citogenético/métodos , Análisis Citogenético/veterinaria , Orden Génico/genética , Humanos , Células Híbridas , Mapeo de Híbrido por Radiación/métodos , Mapeo de Híbrido por Radiación/veterinaria , Translocación GenéticaRESUMEN
There is incredible morphological and behavioral diversity among the hundreds of breeds of the domestic dog, CANIS FAMILIARIS. Many of these breeds have come into existence within the last few hundred years. While there are obvious phenotypic differences among breeds, there is marked interbreed genetic homogeneity. Thus, study of canine genetics and genomics is of importance to comparative genomics, evolutionary biology and study of human hereditary diseases. The most recent version of the map of the canine genome is comprised of 3,270 markers mapped to 3,021 unique positions with an average intermarker distance of approximately 1 Mb. The markers include approximately 1,600 microsatellite markers, about 1,000 gene-based markers, and almost 700 bacterial artificial chromosome-end markers. Importantly, integration of radiation hybrid and linkage maps has greatly enhanced the utility of the map. Additionally, mapping the genome has led directly to characterization of microsatellite markers ideal for whole genome linkage scans. Thus, workers are now able to exploit the canine genome for a wide variety of genetic studies. Finally, the decision to sequence the canine genome highlights the dog's evolutionary and physiologic position between the mouse and human and its importance as a model for study of mammalian genetics and human hereditary diseases.
Asunto(s)
Perros/genética , Genoma , Animales , Evolución Molecular , HumanosRESUMEN
Carbonic anhydrase II (CAII) plays an important role in the acid-base homeostasis of the body and its deficiency results in renal tubular acidosis. In order to identify the regulatory regions in the CAII gene for the future development of kidney-targeted gene therapy, we investigated the 5' region of the gene for its promoter activity. Deletion constructs with various lengths of the 5' flanking region of the human CAII promoter were ligated to the CAT reporter gene and lipofected in primary cultures of mouse proximal renal tubular cells and in cells of the established porcine proximal tubular cell line, LLC-PK1. The CAT activity was measured 48 hours after gene transfection. The -12000/CAT and -1300/CAT constructs expressed the highest CAT activity in both types of renal tubular cells (143- and 180-fold increase, respectively, in mouse proximal tubular cells; 50- and 70-fold increase, respectively, in LLC-PK1 cells) but not the -420/CAT, -270/CAT, or -180/CAT constructs (9, 12, and 9% of that of -1300/CAT construct, respectively, in mouse proximal tubular cells and, 23, 9, and 8%, respectively, in LLC-PK1 cells, all p <0.01 vs. -1300/CAT construct). No cytotoxicity was detected in the transfected cells. A computer search identified multiple putative transcription factor binding elements including Ap1 and Ap2 binding elements, which are present in the -1300/CAT construct but not in the shorter constructs. In conclusion, we demonstrate that the human CAII 5' sequence of proximal 1.3 kb contains strong promoter sequence(s) for renal tubular cells.
Asunto(s)
Anhidrasas Carbónicas/genética , Túbulos Renales Proximales/enzimología , Regiones Promotoras Genéticas/fisiología , Secuencias Reguladoras de Ácidos Nucleicos , Animales , Secuencia de Bases , Supervivencia Celular/fisiología , Células Cultivadas , Cloranfenicol O-Acetiltransferasa/genética , Cloranfenicol O-Acetiltransferasa/metabolismo , Regulación Enzimológica de la Expresión Génica , Genes Reporteros , Vectores Genéticos , Humanos , Túbulos Renales Proximales/fisiología , Ratones , Porcinos , TransfecciónRESUMEN
Von Willebrand's Disease (vWD) in the Scottish Terrier breed is a serious, often fatal, hereditary bleeding disorder. Elimination of the mutated gene by selective breeding is an important goal for the health of this breed. Although the standard protein-based tests are accurate for identification of affected Scottish Terriers, they are not reliable for the identification of carriers of the mutant gene unless multiple replicate assays are performed. A simple, highly accurate test for carriers of the disease is needed so that veterinarians can counsel clients on which animals to use in their breeding programs. The complete coding region of von Willebrand factor (vWF) complementary DNA (cDNA) was sequenced from an affected animal, and a single base deletion in the codon for amino acid 85 of the prepro-vWF cDNA that leads to Scottish Terrier vWD was identified. A highly accurate polymerase chain reaction assay was developed that can distinguish homozygous normal animals from those that are homozygous affected or heterozygous. In a voluntary survey of 87 animals provided by Scottish Terrier owners, 15 were carriers and 4 were affected with vWD, 2 of which had previously been shown to have undetectable vWF. The determination of the complete canine vWF cDNA sequence should facilitate the identification of additional vWD alleles in other breeds and other species.
Asunto(s)
ADN Complementario/genética , Enfermedades de los Perros/genética , Eliminación de Gen , Enfermedades de von Willebrand/veterinaria , Animales , Secuencia de Bases , ADN Complementario/análisis , Perros , Femenino , Pruebas Genéticas , Datos de Secuencia Molecular , Mutación , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Análisis de Secuencia de ADN , Enfermedades de von Willebrand/genéticaRESUMEN
The gammaherpesvirus bovine herpesvirus-4 (BHV-4) has been isolated from a wide variety of animals, including lions and domestic cats. Although BHV-4 antibodies have been detected in normal cats and cats with urinary disorders, the epidemiology and pathogenic role of BHV-4 in cats is unknown. The purpose of this study was to determine the prevalence of BHV-4 antibodies and viral nucleic acid in a population of free-roaming cats. Plasma and peripheral blood leukocyte samples were collected from 52 male and 52 female free-roaming cats impounded at a regional animal control facility in Central Michigan. Plasma concentrations of BHV-4 antibodies were measured with an indirect fluorescent antibody test. Peripheral blood leukocyte DNA was isolated, and a 2-stage polymerase chain reaction with heminested primers delineating a conserved portion of the BHV-4 glycoprotein B gene homologue was used to amplify BHV-4-specific DNA sequences. BHV-4 antibodies were detected in 38 (73%) male and 23 (44%) female cats. Seropositive cats were significantly more likely to be male than female (odds ratio = 3.22; P = .007). Cell-associated viremia was detected in 17 (33%) male and 11 (21%) female cats. Of the 61 seropositive cats, 23 (38%) had a detectable viremia; only 5 (12%) seronegative cats had detectable viremia. Seropositive cats were significantly more likely to be viremic than seronegative cats (OR = 4.30: P = .009). Our results suggest that BHV-4 infection may be more widespread in certain cat populations than previously reported. Furthermore, many cats seropositive for BHV-4 antibodies have a concurrent cell-associated viremia.
Asunto(s)
Anticuerpos Antivirales/sangre , Enfermedades de los Gatos/epidemiología , Gammaherpesvirinae/inmunología , Infecciones por Herpesviridae/veterinaria , Animales , Enfermedades de los Gatos/virología , Gatos , Cartilla de ADN/química , ADN Viral/sangre , ADN Viral/química , ADN Viral/aislamiento & purificación , Electroforesis en Gel de Agar/veterinaria , Electroforesis en Gel de Poliacrilamida/veterinaria , Femenino , Técnica del Anticuerpo Fluorescente Indirecta/veterinaria , Gammaherpesvirinae/genética , Gammaherpesvirinae/aislamiento & purificación , Infecciones por Herpesviridae/sangre , Infecciones por Herpesviridae/epidemiología , Infecciones por Herpesviridae/virología , Masculino , Michigan/epidemiología , Reacción en Cadena de la Polimerasa/veterinaria , Análisis de Secuencia de ADN , Estudios Seroepidemiológicos , Viremia/veterinariaRESUMEN
Interleukin-1 is considered a central mediator of cartilage loss in osteoarthritis in several species, however an equine recombinant form of this cytokine is not readily available for in vitro use in equine osteoarthritis research. Equine recombinant interleukin-1beta was cloned and expressed and its effects on the expression and activity of selected chondrocytic proteins implicated in cartilage matrix degradation were characterized. Reverse transcriptase polymerase chain reaction methods were used to amplify the entire coding region of the equine IL-1beta mRNA, which was cloned into an expression vector, expressed in E. coli, and purified using a Ni2+ chromatographic method. The effects of the recombinant peptide on chondrocyte gene expression were determined by Northern blotting using RNA from equine chondrocyte cultures hybridized to probes for matrix metalloproteinases (MMP 1, MMP 3, MMP 13), tissue inhibitor of matrix metalloproteinases 1 (TIMP 1) and cyclooxygenase 2 (COX 2). Effects on selected mediators of cartilage degradation (nitrite concentrations and MMP activity) were determined using conditioned medium from reIL-1beta-treated equine cartilage explant cultures. A recombinant peptide of approximately 21 kd was obtained. Northern blotting analyses revealed a marked up-regulation of expression of all MMPs, TIMP 1, and COX 2 in mRNA from treated chondrocytes. Furthermore, cartilage explants exposed to reIL-1beta had augmented collagenase/gelatinase and stromelysin activities as well as increased concentration of nitrite in conditioned media. The development of a biologically active, species-specific IL-1beta provides a valuable tool in the study of osteoarthritis pathophysiology and its treatment in horses.
Asunto(s)
Cartílago Articular/efectos de los fármacos , Condrocitos/efectos de los fármacos , Enfermedades de los Caballos/fisiopatología , Interleucina-1/fisiología , Osteoartritis/veterinaria , Animales , Northern Blotting/veterinaria , Cartílago Articular/enzimología , Cartílago Articular/patología , Células Cultivadas , Condrocitos/enzimología , Ciclooxigenasa 2 , Regulación Enzimológica de la Expresión Génica , Caballos , Interleucina-1/farmacología , Isoenzimas/genética , Isoenzimas/metabolismo , Metaloproteinasas de la Matriz/genética , Metaloproteinasas de la Matriz/metabolismo , Osteoartritis/fisiopatología , Prostaglandina-Endoperóxido Sintasas/genética , Prostaglandina-Endoperóxido Sintasas/metabolismo , ARN Mensajero/metabolismo , Proteínas Recombinantes/metabolismo , Proteínas Recombinantes/farmacología , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/veterinaria , Inhibidor Tisular de Metaloproteinasa-1/genética , Inhibidor Tisular de Metaloproteinasa-1/metabolismoRESUMEN
OBJECTIVE: To develop new and improved tests to detect alleles at codons 136 and 171 of the ovine prion protein locus and to evaluate the frequency of these alleles. ANIMALS: 159 Suffolk sheep belonging to 3 flocks. PROCEDURE: Polymerase chain reaction (PCR) analysis that contained diagnostic restriction site variation for each allele were developed for the relevant gene regions. Alleles were determined by analyzing DNA isolated from buccal swab specimens or blood samples. RESULTS: At codon 136, frequencies of the alanine and valine alleles were found to be 97 and 3%, respectively. At codon 171, frequencies of the glutamine, arginine, and histidine alleles were found to be 57, 41, and 2%, respectively. CONCLUSIONS: Little variation was detected in codon 136, whereas noteworthy variation was found in codon 171; > 40% of the alleles at this locus coded for glutamine. Because the glutamine allele at codon 171 confers susceptibility to scrapie, reduction of its frequency is of importance to management of sheep flocks. CLINICAL RELEVANCE: Genotyping of sheep, using the tests reported here, should facilitate selective breeding programs designed to decrease the risk of scrapie.
Asunto(s)
Codón/genética , Reacción en Cadena de la Polimerasa/veterinaria , Enfermedades por Prión/veterinaria , Priones/genética , Enfermedades de las Ovejas/genética , Alelos , Animales , ADN/sangre , Cartilla de ADN/química , Enzimas de Restricción del ADN/química , Electroforesis en Gel de Agar/veterinaria , Mucosa Bucal/química , Reacción en Cadena de la Polimerasa/métodos , Enfermedades por Prión/diagnóstico , Enfermedades por Prión/genética , Análisis de Secuencia de ADN , Ovinos , Enfermedades de las Ovejas/diagnósticoRESUMEN
OBJECTIVE: To identify a DNA marker for the copper toxicosis (CT) locus in Bedlington Terriers (BT). ANIMALS: 77 BT, of which 25 were affected. Diagnosis of affected or unaffected with CT was made in all cases by quantitative copper determinations on liver biopsy samples by use of established criteria. PROCEDURE: BT pedigrees segregating for CT were identified. Linkage studies were carried out using polymorphic microsatellite markers developed for the canine genome in these pedigrees. DNA was isolated from blood samples of pedigree members. Polymerase chain reaction was used to amplify and type alleles at 213 microsatellite loci in each dog, and findings were subjected to linkage analysis. RESULTS: One microsatellite marker was identified to be closely linked to CT with logarithm of odds score of 5.96 at a recombination fraction of zero. CONCLUSIONS: Using the linked marker, it has become possible to distinguish affected, homozygous normal, and carrier dogs in some BT pedigrees. CLINICAL RELEVANCE: In informative pedigrees where the marker is variable in the parents, it is possible to identify which dogs will require anticopper therapy and provide breeders with sound scientific advice about breeding strategies.
Asunto(s)
Cobre/toxicidad , ADN Satélite/genética , Enfermedades de los Perros/genética , Enfermedades Genéticas Congénitas/veterinaria , Ligamiento Genético , Repeticiones de Microsatélite , Alelos , Animales , Cobre/metabolismo , Enfermedades de los Perros/metabolismo , Perros , Femenino , Genes Recesivos , Enfermedades Genéticas Congénitas/metabolismo , Marcadores Genéticos , Homocigoto , Masculino , Linaje , Reacción en Cadena de la Polimerasa/veterinariaRESUMEN
The exact cause of hematuria, dysuria, and urethral obstruction remains unknown in a large percentage of naturally occurring cases of feline lower urinary tract disease (FLUTD). One attractive hypothesis implicates viruses as the cause of some idiopathic forms of FLUTD; supporting this hypothesis is the fact that a gamma herpesvirus, a calicivirus, and a retrovirus have been isolated from urine and tissues obtained from cats with this type of disease. Although the clinical course and laboratory findings of cats with idiopathic FLUTD are suggestive of an infectious cause, the question of whether viruses have a pathologic role in some forms of naturally acquired FLUTDs has not been completely answered.
Asunto(s)
Enfermedades de los Gatos/virología , Infecciones Urinarias/veterinaria , Virosis/veterinaria , Animales , Antivirales/uso terapéutico , Infecciones por Caliciviridae/diagnóstico , Infecciones por Caliciviridae/veterinaria , Infecciones por Caliciviridae/virología , Calicivirus Felino , Enfermedades de los Gatos/diagnóstico , Enfermedades de los Gatos/tratamiento farmacológico , Gatos , Herpesviridae , Infecciones por Herpesviridae/diagnóstico , Infecciones por Herpesviridae/veterinaria , Infecciones por Herpesviridae/virología , Infecciones por Retroviridae/diagnóstico , Infecciones por Retroviridae/veterinaria , Infecciones por Retroviridae/virología , Spumavirus , Sistema Urinario/efectos de los fármacos , Sistema Urinario/virología , Infecciones Urinarias/diagnóstico , Infecciones Urinarias/virología , Virosis/diagnóstico , Virosis/virologíaRESUMEN
BACKGROUND: Selective intestinal cobalamin malabsorption with mild proteinuria (Imerslund-Gräsbeck syndrome; I-GS), is an autosomal recessive disorder of dogs caused by mutations in AMN or CUBN that disrupt cubam function and which can present as a medical emergency. OBJECTIVES: To describe the clinical, metabolic, and genetic bases of I-GS in Beagles. ANIMALS: Four cobalamin-deficient and 43 clinically normal Beagles and 5 dogs of other breeds. METHODS: Clinical description and candidate gene genetic study. Urinary organic acid and protein excretion were determined by gas-chromatography and SDS-PAGE, respectively. Renal cubilin protein expression was assessed on immunoblots. Mutation discovery was carried out by PCR amplification and DNA sequencing of exons with flanking splice sites and cDNA of CUBN and AMN. Genotyping was performed by restriction enzyme digestion of PCR amplicons. RESULTS: Juvenile-affected Beagles exhibited failure to thrive, dyshematopoiesis with neutropenia, serum cobalamin deficiency, methylmalonic aciduria, hyperammonemia, and proteinuria. Affected dogs' kidneys lacked detectable cubilin protein. All affected dogs were homozygous for a single-base deletion in CUBN exon 8 (CUBN c.786delC), predicting a translational frameshift, and the 2 parents tested were heterozygous. CONCLUSIONS: The CUBN mutation in juvenile I-GS Beagles causes a more severe cobalamin malabsorption than in Border Collies with a different CUBN defect, but is similar to I-GS caused by AMN mutations in Giant Schnauzers and Australian Shepherds. Awareness of the disorder and breed predispositions to I-GS is crucial to precisely diagnose and promptly treat hereditary cobalamin malabsorption and to prevent disease in those dogs at risk in future generations.