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Biomedical subjects

G Diane Shelton

Publications and source records attributed to G Diane Shelton.

At least 19 recordsLinked to original sources
PubMed2006

Identification of a canine model of pyruvate dehydrogenase phosphatase 1 deficiency.

Exercise intolerance syndromes are well known to be associated with inborn errors of metabolism affecting glycolysis (phosphorylase and phosphofructokinase deficiency) and fatty acid oxidation (palmitoyl carnitine transferase deficiency). We have identified a canine model for profound exercise intolerance caused by a deficit in PDP1 (EC 3.1.3.43), the phosphatase enzyme that activates the pyruvate dehydrogenase complex (PDHc). The Clumber spaniel breed was originated in 1760 by the Duc de Noailles, as a hunting dog with a gentle temperament suitable for the 'elderly gentleman'. Here we report that 20% of the current Clumber and Sussex spaniel population are carriers for a null mutation in PDP1, and that homozygosity produces severe exercise intolerance. Human pyruvate dehydrogenase phosphatase deficiency was recently characterized at the molecular level. However, the nature of the human mutation (loss of a single amino acid altering PDP1 activity) made it impossible to discern the role of the second phosphatase isoform, PDP2, in the deficient phenotype. Here we show that the null mutation in dogs provides a valuable animal model with which to study the effects of dysregulation of the PDHc. Knowledge of the molecular defect has allowed for the institution of a rapid restriction enzyme test for the canine mutation that will allow for selective breeding and has led to a suggested dietary therapy for affected dogs that has proven to be beneficial. Pharmacological and genetic therapies for PDP1 deficiency can now be investigated and the role of PDP2 can be fully characterized.

Animals↗
PubMed2006

Sarcolemma-specific autoantibodies in canine inflammatory myopathy.

Inflammatory myopathies (IM) are relatively common in dogs with an increased incidence in the Boxer and Newfoundland breeds. Here, we show that a high proportion of affected Boxers and Newfoundlands have circulating autoantibodies against unknown sarcolemma antigens, that are muscle-specific but not species specific. We further show that the autoantigen can be extracted from muscle membranes with non-ionic detergent, and that such detergent extracts can be used in a sensitive ELISA for detection and quantitation of antibodies. The relatively high incidence of IM with autoantibodies in selected breeds of dogs indicates a genetic predisposition for a particular form of IM. In these breeds, this form of IM could be diagnosed and monitored with a simple serum assay.

Animals↗
PubMed2006

Immunopathogenic pathways in canine inflammatory myopathies resemble human myositis.

Progress in the treatment of inflammatory myopathies is impeded by the lack of suitable animal models. Inflammatory myopathies occur spontaneously in the dog, are a heterogeneous group of disorders, and are more common than in humans. Clinical signs of weakness and muscle atrophy are reliably present, and there are histological and immunohistological similarities to forms of human myositis. In this study, microarray technology followed by quantitative real-time PCR and immunohistochemistry on muscle biopsy sections was used to investigate gene expression in cases of canine inflammatory myopathies. Several genes involved with innate and adaptive immunity were highly upregulated including those that participate in macrophage and dendritic cell activation and migration, and antigen processing and presentation. Other genes including those that participate in B cell growth, development, migration and activation, immunoglobulin genes, genes in pro-inflammatory and anti-inflammatory pathways, and genes involved with tissue remodeling were upregulated. In previous reports utilizing microarray technology in human myositis, there was activation of similar pathways involved in the immune response. This study strengthens the argument that forms of canine myositis may be important animal models of human myositis and suggests useful biomarkers for therapeutic response using the dog in pre-clinical trials.

Animals↗
PubMed2006

Neurological disease in wild loggerhead sea turtles Caretta caretta.

Beginning in October 2000, subadult loggerhead sea turtles Caretta caretta showing clinical signs of a neurological disorder were found in waters off south Florida, USA. Histopathology indicated generalized and neurologic spirorchiidiasis. In loggerhead sea turtles (LST) with neurospirorchiidiasis, adult trematodes were found in the meninges of the brain and spinal cord of 7 and 3 affected turtles respectively, and multiple encephalic intravascular or perivascular eggs were associated with granulomatous or mixed leukocytic inflammation, vasculitis, edema, axonal degeneration and occasional necrosis. Adult spirorchiids were dissected from meningeal vessels of 2 of 11 LST brains and 1 of 10 spinal cords and were identified as Neospirorchis sp. Affected LST were evaluated for brevetoxins, ciguatoxins, saxitoxins, domoic acid and palytoxin. While tissues from 7 of 20 LST tested positive for brevetoxins, the levels were not considered to be in a range causing acute toxicosis. No known natural (algal blooms) or anthropogenic (pollutant spills) stressors co-occurred with the turtle mortality. While heavy metal toxicosis and organophosphate toxicosis were also investigated as possible causes, there was no evidence for their involvement. We speculate that the clinical signs and pathologic changes seen in the affected LST resulted from combined heavy spirorchiid parasitism and possible chronic exposure to a novel toxin present in the diet of LST.

Animals↗
PubMed2006

Congenital myopathy with abundant nemaline rods in a cat.

Nemaline myopathy is associated with rod-shaped structures in muscle fibers. At least seven distinct clinical forms have been described in humans and mutations have been identified in five different thin-filament genes. Only a few cases of spontaneously occurring nemaline myopathy have been reported in animals and include an adult-onset form in a family of cats and an early-onset form in a dog. Here, we describe a 2-year-old male, neutered, domestic shorthaired cat that was referred to the Veterinary Medical Teaching Hospital, University of California-Davis, for evaluation of chronic, progressive weakness, and fine tremors. Neurologic deficits were restricted to the neuromuscular system. Electromyography showed mild to moderate diffuse spontaneous activity. Although rod bodies were prominent on light and electron microscopic evaluation of biopsies from several muscles, sarcoplasmic accumulations of dystrophin, desmin, and spectrin also were identified by immunohistochemistry. These findings may represent the occurrence of rod bodies in conjunction with a protein-aggregate myopathy.

Animals↗
PubMed2006

BAG3 deficiency results in fulminant myopathy and early lethality.

Bcl-2-associated athanogene 3 (BAG3) is a member of a conserved family of cyto-protective proteins that bind to and regulate Hsp70 family molecular chaperones. Here, we show that BAG3 is prominently expressed in striated muscle and colocalizes with Z-disks. Mice with homozygous disruption of the bag3 gene developed normally but deteriorated postnatally with stunted growth evident by 1 to 2 weeks of age and death by 4 weeks. BAG3-deficient animals developed a fulminant myopathy characterized by noninflammatory myofibrillar degeneration with apoptotic features. Knockdown of bag3 expression in cultured C2C12 myoblasts increased apoptosis on induction of differentiation, suggesting a need for bag3 for maintenance of myotube survival and confirming a cell autonomous role for bag3 in muscle. We conclude that although BAG3 is not required for muscle development, this co-chaperone appears to be critically important for maintenance of mature skeletal muscle.

Adaptor Proteins, Signal Transducing↗
PubMed2006

Inherited myopathy in a young Great Dane.

A 3-year-old, male Great Dane was evaluated for an 18-month history of progressive weakness. Histologic evaluation of muscle biopsies revealed distinct cytoarchitectural changes that were indistinguishable from the central "core-like" structures previously described as central core myopathy in this breed. Clinical features of this inherited myopathy are described.

Animals↗
PubMed2005

Myokymia and neuromyotonia in a cat.

A 6-year-old spayed female domestic shorthair cat was examined because of a 2-week history of rhythmic muscle movements. Physical examination revealed thoracic limb rigidity, contracture of the carpi, generalized muscle atrophy, and rhythmic rippling of the muscles of all 4 limbs. Results of a CBC and serum biochemistry profile were unremarkable other than high creatine kinase activity. Electromyography revealed unique high-frequency discharges, including rhythmic bursts of single motor unit potentials appearing as doublets (myokymia) and more prolonged bursts of nonrhythmic motor unit potentials with characteristic waning amplitudes (neuromyotonia). Histologic examination of muscle biopsy specimens revealed noninflammatory necrotizing myopathy with regeneration. The cat did not respond to treatment with carbamazepine or prednisone but improved rapidly after treatment with phenytoin was initiated. Six months after initial examination, electromyography revealed a substantial decrease in the amount of spontaneous activity in previously affected muscles. However, the myokymic and neuromyotonic discharges were still present, albeit with a substantial decrease in frequency.

Animals↗
PubMed2005

Sensory and motor neuropathy in a Border Collie.

A 5-month-old female Border Collie was evaluated because of progressive hind limb ataxia. The predominant clinical findings suggested a sensory neuropathy. Sensory nerve conduction velocity was absent in the tibial, common peroneal, and radial nerves and was decreased in the ulnar nerve; motor nerve conduction velocity was decreased in the tibial, common peroneal, and ulnar nerves. Histologic examination of nerve biopsy specimens revealed considerable nerve fiber depletion; some tissue sections had myelin ovoids, foamy macrophages, and axonal degeneration in remaining fibers. Marked depletion of most myelinated fibers within the peroneal nerve (a mixed sensory and motor nerve) supported the electrodiagnostic findings indicative of sensorimotor neuropathy. Progressive deterioration in motor function occurred over the following 19 months until the dog was euthanatized. A hereditary link was not established, but a littermate was similarly affected. The hereditary characteristic of this disease requires further investigation.

Animals↗
PubMed2005

ADAM12 overexpression does not improve outcome in mice with laminin alpha2-deficient muscular dystrophy.

We have recently shown that overexpression of ADAM12 results in increased muscle regeneration and significantly reduced pathology in mdx, dystrophin deficient mice. In the present study, we tested the effect of overexpressing ADAM12 in dy(W) laminin-deficient mice. dy mice have a very severe clinical phenotype and would be expected to benefit greatly from enhanced regeneration. We found that dy(W) mice overexpressing ADAM12 indeed have increased muscle regeneration, as evidenced by increased numbers of muscle fibers expressing fetal myosin. However, overexpression of ADAM12 had no significant effect on overall health, as evidenced by body weight, and did not improve muscle pathology.

ADAM Proteins↗
PubMed2005

Canine spongiform leukoencephalomyelopathy is associated with a missense mutation in cytochrome b.

Two families of dogs (Australian cattle dogs and Shetland sheepdogs) with an inherited "spongiform leukoencephalomyelopathy" were identified, with widespread vacuolation of white matter of the brain and spinal cord. Affected dogs of both breeds developed tremors at 2-9 weeks of age followed by progressive neurological worsening with ataxia, paresis, paralysis, spasticity, and cranial nerve dysfunction. The modes of inheritance of both families were most likely maternal. The cerebrospinal fluid (CSF) analysis showed elevated ratio of 3-OH butyrate to acetoacetic acid. Mitochondrial DNA sequencing showed a G to A transition at 14,474 nt (G14474A, GenBank accession no. NC002008 ) that results in an amino acid change of valine-98 to methionine (V98M) of mitochondrial encoded cytochrome b. Western blot analysis showed increased levels of core I and core II but decreased level of cytochrome c1 of the complex III and cytochrome c oxidase of the complex IV of the respiratory chain.

3-Hydroxybutyric Acid↗
PubMed2005

Dysphagia as the primary clinical abnormality in two dogs with inflammatory myopathy.

Two adult Boxers were evaluated because of chronic dysphagia of several years' duration. Serum creatine kinase activity was high in both dogs, but other hematologic or serum biochemical abnormalities were not detected. Esophagraphy revealed abnormalities of the cricopharyngeal phase of swallowing in both dogs, and electromyography of the pharyngeal and laryngeal muscles revealed complex repetitive discharges, positive sharp waves, and fibrillation potentials characteristic of primary myopathy or neuropathy. Because of the severity of their condition, both dogs were euthanatized. Histologically, mixed-cell infiltrates were seen in sections of the masseter and thyropharyngeal muscles. Results of indirect immunofluorescence staining for proteins associated with dystrophic myopathy were unremarkable, except for decreased staining for integrin alpha7. A diagnosis of chronic inflammatory myopathy was made. The clinical importance of reduced staining for integrin alpha7 could not be determined but was considered to be a result of the myopathy.

Animals↗
PubMed2005

Canine and feline models of human inherited muscle diseases.

Animal models are of immense importance for studying mechanisms of disease and testing new therapies, and rodents have been used extensively in the field of neuromuscular disorders. Mice and rats can be genetically manipulated to over-express or not express genes that are important to muscle function, and these animals can be available in large numbers for analysis. Other species, such as cats and dogs, cannot be manipulated in the same ways or be used in large numbers, but they have spontaneously occurring muscle diseases with clinical presentations more closely resembling those of the human disorders. Therefore, cats and dogs may become valuable as intermediate disease models. This review focuses on canine and feline models of human inherited muscle diseases with comparisons to rodent models and an emphasis on the muscular dystrophies.

Animals↗
PubMed2005

Expanded repeat in canine epilepsy.

Epilepsy afflicts 1% of humans and 5% of dogs. We report a canine epilepsy mutation and evidence for the existence of repeat-expansion disease outside humans. A canid-specific unstable dodecamer repeat in the Epm2b (Nhlrc1) gene recurrently expands, causing a fatal epilepsy and contributing to the high incidence of canine epilepsy. Tracing the repeat origins revealed two successive events, starting 50 million years ago, unique to canid evolution. A genetic test, presented here, will allow carrier and presymptomatic diagnosis and disease eradication. Clinicopathologic characterization establishes affected animals as a model for Lafora disease, the most severe teenage-onset human epilepsy.

Alleles↗
PubMed2005

Inherited motor neuron disease in domestic cats: a model of spinal muscular atrophy.

Juvenile-onset spinal muscular atrophy was observed in an extended family of purebred domestic cats as a fully penetrant, simple autosomal recessive trait. Affected kittens exhibited tremor, proximal muscle weakness, and muscle atrophy beginning at ~4 mo of age. Apparent loss of function was rapid initially but progressed slowly after 7-8 mo of age, and variably disabled cats lived for at least 8 y. Electromyography and microscopic examination of muscle and nerve biopsies were consistent with denervation atrophy as a result of a central lesion. There was astrogliosis and dramatic loss of motor neurons in ventral but not dorsal horn gray matter of spinal cord and loss of axons in ventral horn nerve roots. These phenotypic findings were similar to mild forms (type III) of spinal muscular atrophy in humans caused by survival of motor neuron mutations, but molecular analysis excluded feline survival of motor neuron as the disease gene in this family. A breeding colony has been established for further investigation of this naturally occurring large-animal model of inherited motor neuron disease.

Age Factors↗
PubMed2005

Muscle regeneration, inflammation, and connective tissue expansion in canine inflammatory myopathy.

Inflammatory myopathies (IMs) are relatively common in dogs, and canine IMs have many similarities to human IMs. The aim of this work was to analyze aspects of the pathogenesis of canine IM with an ultimate goal of establishing canine IM as a model for human IM. Muscle biopsies from 16 dogs with a histological diagnosis of IM were analyzed to determine degree of muscle regeneration, presence of eosinophils, expression of selected cytokines and chemokines, and extent of fibrosis. Regeneration, as shown by staining for developmental myosin heavy chain, was more extensive than evidenced with hematoxylin-eosin staining in most cases of canine IM. Expression of mRNA encoding transforming growth factor-beta (TGF-beta) and eotaxin 3 was upregulated in all cases evaluated. Eosinophils were abundant in most cases, and the connective tissue was variably expanded, as demonstrated by the distribution of the ubiquitous extracellular matrix proteins collagen VI and fibrillin. The extensive regeneration demonstrates that muscle may survive this adverse environment if inflammation and fibrosis can be stopped or reduced.

Animals↗
PubMed2005

Elimination of myostatin does not combat muscular dystrophy in dy mice but increases postnatal lethality.

Myostatin is a TGF-beta family member and a negative regulator of skeletal muscle growth. It has been proposed that reduction or elimination of myostatin could be a treatment for degenerative muscle diseases such as muscular dystrophy. Laminin-deficient congenital muscular dystrophy is one of the most severe forms of muscular dystrophy. To test the possibility of ameliorating the dystrophic phenotype in laminin deficiency by eliminating myostatin, we crossed dy(W) laminin alpha2-deficient and myostatin null mice. The resulting double-deficient dy(W)/dy(W);Mstn(-/-) mice had a severe clinical phenotype similar to that of dy(W)/dy(W) mice, even though muscle regeneration was increased. Degeneration and inflammation of muscle were not alleviated. The pre-weaning mortality of dy(W)/dy(W);Mstn(-/-) mice was increased compared to dy(W)/dy(W), most likely due to significantly less brown and white fat in the absence of myostatin, and postweaning mortality was not significantly improved. These results show that eliminating myostatin in laminin-deficiency promotes muscle formation, but at the expense of fat formation, and does not reduce muscle pathology. Any future therapy based on myostatin may have undesirable side effects.

Adipocytes↗