In vitro and in vivo gene therapy with CMV vector-mediated presumed dog ��-nerve growth factor in pyridoxine-induced neuropathy dogs

Due to the therapeutic potential of gene therapy for neuronal injury, many studies of neurotrophic factors, vectors, and animal models have been performed. The presumed dog β-nerve growth factor (pdβ-NGF) was generated and cloned and its expression was confirmed in CHO cells. The recombinant pdβ-NGF protein reacted with a human β-NGF antibody and showed bioactivity in PC12 cells. The pdβ-NGF was shown to have similar bioactivity to the dog β-NGF. The recombinant pdβ-NGF plasmid was administrated into the intrathecal space in the gene therapy group. Twenty-four hours after the vector inoculation, the gene therapy group and the positive control group were intoxicated with excess pyridoxine for seven days. Each morning throughout the test period, the dogs'' body weight was taken and postural reaction assessments were made. Electrophysiological recordings were performed twice, once before the experiment and once after the test period. After the experimental period, histological analysis was performed. Dogs in the gene therapy group had no weight change and were normal in postural reaction assessments. Electrophysiological recordings were also normal for the gene therapy group. Histological analysis showed that neither the axons nor the myelin of the dorsal funiculus of L₄ were severely damaged in the gene therapy group. In addition, the dorsal root ganglia of L₄ and the peripheral nerves (sciatic nerve) did not experience severe degenerative changes in the gene therapy group. This study is the first to show the protective effect of NGF gene therapy in a dog model.     

Distinctive histopathologic features of canine optic nerve hypoplasia and aplasia: a retrospective review of 13 cases

Canine optic nerve hypoplasia (ONH) and aplasia (ONA) are significant neuro-ophthalmologic disorders that have been reported in several species. The purpose of this study was to describe the distinctive histopathologic features of ONH and ONA in canine patients identified from a collection of 20 000 ocular submissions at the comparative ocular pathology laboratory of Wisconsin from 1989 to 2006. The following information about ONH and ONA cases was collected: signalment, and clinical and gross findings, including unilateral vs. bilateral involvement. Microscopic evaluation was performed, with attention to optic nerve malformation, retinal ganglion cell (RGC) and nerve fiber layer (NFL) loss, and retinal disorganization. The distribution of retinal vasculature was recorded and a search for unusual findings of ONH and ONA was performed. Information and histologic documentation was available for 13 cases. Eight cases of ONH and five cases of ONA were identified. The average group age was 20.2 months and 16.1 months, respectively. The most common breed was the Shih Tzu (3/13). ONH usually presented bilaterally (7/8); all ONA cases presented as a unilateral disease (5/5). The morphologic findings in the optic nerve (ON) in ONH included variable degrees of ON hypoplasia and gliosis, as well as ectopic vestigial ON remnants within orbital nerves and connective tissues. The NFL was detected in the majority of the ONH cases; however, RGCs were rare or absent. Mild retinal disorganization was seen occasionally. Most cases of ONH were associated with regional peripheral retinal blood vessel extension into the vitreous, leaving the peripheral retina avascular. In ONA cases the retinal blood vessels, NFL and RGCs were totally absent and retinal disorganization was severe. Distinctive microscopic features encountered in ONA included anterior segment dysgenesis in some cases. The retina in these cases was stretched across the posterior lens capsule, never making contact with the posterior pole of the globe. The current study reviews the human and veterinary literature pertaining to ONH and ONA, compares ONH and ONA in dogs, and presents related ophthalmic histopathologic findings that have not been reported previously.     

Blindness in a wild American black bear cub (Ursus americanus)

An approximately six‐month‐old wild American black bear (Ursus americanus) was found wandering in Saskatchewan and was presented to the Veterinary Medical Centre of the Western College of Veterinary Medicine for apparent blindness. Clinical examination confirmed an inability to navigate a photopic maze, bilateral tapetal hyper‐reflectivity, fundi devoid of retinal vessels, and small pale optic nerve papillae. Single‐flash electroretinography revealed A and B‐wave amplitudes of approximately 40 and 140 microvolts, respectively, in both eyes. Histologic abnormalities included bilateral optic papillary mineralization and bilateral segmental optic nerve degeneration, with occasional intralesional lymphocytes confirmed with immunohistochemistry for CD3+. There was also bilateral multifocal retinal dysplasia, gliosis, lymphocytic retinitis, a complete lack of retinal blood vessels, an intravitreal vascular membrane, and a mild lymphocytic–plasmacytic uveitis with small pre‐iridal cellular membranes. The presence of a positive ERG in a blind bear with numerous retinal ganglion cells and degenerative changes in the optic nerve are most consistent with vision loss due to optic nerve injury, which given the young age of the bear likely occurred during ocular development. The presence of ocular inflammation suggests this injury resulted from an inflammatory/infectious process. The etiology could not be determined. Hepatic concentrations of vitamin A were within the normal reference range for domestic species. Pan‐herpesvirus PCR and immunohistochemistry for canine distemper virus and Toxoplasma gondii were negative, although this does not rule out these or other infectious etiologies. This represents the first case report of neonatal or congenital ocular abnormalities in an ursid species.     

Effect of renal denervation by radiofrequency catheter ablation on expression of aquaporins in dog kidneys

AIM: To investigate the effect of renal denervation (RDN) by radiofrequency catheter ablation on the expression of aquaporins (AQP) in dog kidneys.METHODS: Adult Chinese Kunming dogs (n=12) were randomly divided into RDN group and control group (6 for each group). The dogs in RDN group underwent bilateral RDN using radiofrequency catheter ablation, and radiofrequency catheter was positioned in bilateral renal artery without ablation in control group. The levels of norepinephrine (NE) and AQP1~3 in the renal tissues were detected 1 month after RDN, and blood pressure (BP) measurements were performed at baseline and 1 month after RDN.RESULTS: The level of NE in RDN group was significantly lower than that in control group (P<0.01). The expression of AQP1~3 in the renal cortex and medulla was lower in RDN group than that in control group. RDN also caused a substantial BP reduction (P<0.05). CONCLUSION: RDN substantially decreases the tissue levels of NE and AQP in dog kidneys, and also decreases BP significantly, which might be involved in the mechanism of BP reduction by RDN. Renal sympathetic nerve plays an excitatory role in the regulation of AQP in the kidney.     

Non-contact mapping and frequency analysis of vagal-mediated atrial fibrillation

AIM: To explore the mechanism of the initiation and maintenance of vagal-mediated atrial fibrillation (AF) by non-contact mapping and frequency analysis of vagal-mediated atrial fibrillation in canine.METHODS: Atrial effective refractory period (AERP) and dispersion of AERP were measured in 8 canine during baseline and bilateral cervical vagal nerve stimulation (CVNS). Left atrium (LA) and right atrium (RA) electrical activity of AF was assessed by non-contact mapping and frequency analysis.RESULTS: Compared with baseline, CVNS attenuated left and right AERP, but only increased the dispersion of left AERP. During CVNS, AF was easily induced and maintained, repetitive organized activations rotated around a preferential route were only found in the LA, and dominant frequencies (DFs) from LA were higher than those of the RA ［(12.5±1.5)Hz vs (9.3±1.2)Hz, P<0.01］. After the cessation of CVNS, DFs of AF decreased in the LA and RA ［(9.2±0.5)Hz vs (8.5±0.6)Hz, P>0.05］, and AF was spontaneously terminated.CONCLUSION: The change of electrophysiological character, difference of activation pattern, and frequency gradient between the LA and RA suggest that the initiation and maintenance of vagal-mediated AF dependent on the LA.     

Genetic analysis of optic nerve head coloboma in the Nova Scotia Duck Tolling Retriever identifies discordance with the NHEJ1 intronic deletion (collie eye anomaly mutation)

Collie eye anomaly (CEA) encompasses a spectrum of different ophthalmic phenotypes from clinically inconsequential choroidal hypoplasia to blindness from coloboma of the optic nerve head (ONH). A previous study found a 7.8‐kb deletion in intron 4 of the NHEJ1 gene to be associated with CEA. A genetic test based on this association is recommended for many breeds, including the Nova Scotia Duck Tolling Retriever (NSDTR). Collection of ONH coloboma‐affected NSDTR showed lack of concordance of the NHEJ1 intronic deletion with ONH coloboma. Using genomewide single nucleotide polymorphism (SNP) genotyping in 7 ONH coloboma‐affected NSDTR cases and 47 unaffected NSDTR controls with no ophthalmic signs, one SNP, located on chromosome 7, demonstrated genomewide significance. However, high genomic inflation may have confounded the results. Therefore, the genomewide association study was repeated using EMMAX to control for population structure in the cohort of 7 cases and 47 controls. However, no regions of the genome were significantly associated with ONH coloboma. These results failed to document significant association with the CEA locus. Due to the complex genetic etiology of ONH coloboma, the NHEJ1 intronic deletion test results should be carefully considered when making breeding decisions. If the goal is to select for visually competent dogs, our data suggest that eye examinations of puppies would be more effective as a guide in selection of breeding pairs than relying solely on currently available genetic tests.