Ouabain, a cardiac glycoside, binds to the Na+-K+i-adenosine triphosphatase (Na+ pump) and prevents active transport of Na+ and K+ across cell membranes. We used [3H]ouabain to quantify the number and affinity of Na+ pumps in skeletal muscle from Quarter Horses with the muscular disorder hyperkalemic periodic paralysis (HYPP). [3H]Ouabain-binding properties of gluteal muscle from clinically normal and affected horses were used to determine whether altered Na+ pump number or affinity could contribute to the pathologic features of muscle in affected horses. Foals and adult horses with HYPP were compared with age-matched clinically normal horses. The number of [3H]ouabain-binding sites in adult gluteal muscle was not different between the 2 types of horses (85.7 +/- 8.9 pmol of [3H]ouabain-binding sites/g [wet muscle weight] in horses with HYPP vs 100.2 +/- 8.8 pmol/g in clinically normal adult horses). Gluteal muscles in HYPP-affected and clinically normal foals also contained a similar number of [3H]ouabain-binding sites (222.3 +/- 21.0 pmol/g vs 225.3 +/- 24.2 pmol/g, respectively). The affinity of these binding sites for ouabain was not different, between adults or foals, in clinically normal or affected horses. Our results indicate that membrane events underlying the periodic episodes of paralysis in horses with HYPP are not attributable to quantitative changes in Na+ pump number or affinity. Our data cannot exclude the possibility that the specific activity of the Na+ pump is altered in muscle from HYPP-affected horses.

OBJECTIVE To sequence exons and splice consensus sites of the dynactin subunit 1 (DCTN1) gene in Leonbergers and Labrador Retrievers with clinical laryngeal paralysis. ANIMALS 5 unrelated Leonbergers with laryngeal paralysis, 2 clinically normal Leonbergers, 7 unrelated Labrador Retrievers with laryngeal paralysis, and 2 clinically normal Labrador Retrievers. PROCEDURES Primers were designed for the entire coding regions of the DCTN1 gene, a noncoding exon at the 5´ end of the gene, and a 900-bp single-nucleotide polymorphism (SNP)-rich region located 17 kb upstream of the DCTN1 gene by use of the CanFam3 assembly of the canine genome sequence. Sequences were generated and compared between clinically normal and affected dogs. The SNPs flanking the DCTN1 gene as well as a previously identified nonsynonymous SNP in exon 32 were genotyped in affected and clinically normal Leonbergers and Labrador Retrievers. RESULTS None of the affected dogs were homozygous for any mutation affecting coding regions or splicing consensus sequences. Of the 16 dogs tested for the missense SNP in exon 32, all were homozygous for the reference allele, except for 2 affected and 1 clinically normal Labrador Retriever and 1 clinically normal Leonberger. The DCTN1 gene sequences (5 dogs) and haplotypes of polymorphic markers surrounding the DCTN1 gene (all dogs) were not consistent with the hypothesis that laryngeal paralysis was associated with inheritance of the same DCTN1 disease-causing allele within all Labrador Retrievers or Leonbergers evaluated. CONCLUSIONS AND CLINICAL RELEVANCE Mutations in the DCTN1 gene did not appear to cause laryngeal paralysis in Leonbergers or Labrador Retrievers.

Effects of ventriculectomy and prosthetic laryngoplasty on upper airway flow mechanics and blood gas tensions in exercising horses with induced left laryngeal hemiplegia were assessed. Five adult horses were trained to stand, trot (4.5 m/s), and gallop (7.2 m/s) on a treadmill (6.38? incline). Inspiratory and expiratory airflows (VImax, VEmax, respectively) were measured using a 15.2-cm diameter pneumotachograph in a face mask. Inspiratory and expiratory transupper airway pressures (PuI, PuE respectively) were determined as pressure differences between barometric pressure and lateral tracheal pressure. Blood collected from exteriorized carotid arteries was analyzed for PaO2, PaCO2, pH, hemoglobin (Hb) content, and HCO3-values. Heart rate (HR) was determined with an HR monitor. Measurements were made with horses standing, trotting, and galloping before left recurrent laryngeal neurectomy (LRLN; base line), 14 days after LRLN, 30 days after ventriculectomy (44 days after LRLN), and 14 days after prosthetic laryngoplasty (58 days after LRLN). Before LRLN (base line), increasing treadmill speed for horses from standing to the trot and gallop progressively increased HR, respiratory frequency, VImax, VEmax, PuI, PuE, Hb, and PaCO2 values and decreased PaO2 pH, and HCO3- values; inspiratory and expiratory impedances were unchanged. After LRLN, inspiratory impedance and PuI were significantly (P < 0.05) increased in horses at the trot and gallop, and PaCO2 was significantly increased in horses at the gallop. The VImax and respiratory frequency were significantly (P < 0.05) decreased in horses at the gallop. Left recurrent laryngeal neurectomy had no effect on PuE VEmax, HR, PaO2, pH, Hb or expiratory impedance values. Ventriculectomy failed to improve upper airway flow mechanics induced by LRLN, whereas prosthetic laryngoplasty restored upper airway flow mechanics to base-line values.

A 5-year-old, castrated male, Maltese was presented with history of acute flaccid paralysis. The dog was presented with sudden loss of muscle tone and involuntary movements of hind limbs. Neurologic examination revealed reduced postural reaction in the bilateral hind limbs. MRI of brain showed moderate hydrocephalus, but other examination results were normal. Based on the characteristic episodes and examination results, canine cataplexy was suspected. Treatment was initiated with clomipramine as cataplexy control. Clinical signs resolved with 3-month medication. This case demonstrates therapeutic diagnosis of cataplexy. To the author’s knowledge, this is the first report of cataplexy treating with clomipramine.

Left laryngeal hemiplegia was induced by resection of the left recurrent laryngeal nerve in 12 dogs. A neuro-muscular pedicle graft formed from the first cervical nerve and sternothyroideus muscle was transplanted after 1 week to the denervated cricoarytenoideus dorsalis muscle in 8 dogs. The remaining 4 dogs served as controls. Left arytenoid abduction was blindly evaluated by laryngoscopy with video photography at time 0, at 1 week, and at 19 weeks in all dogs. At 19 weeks, biopsy specimens of the left cricoarytenoideus dorsalis muscle and the neuromuscular pedicle were taken from 4 of the treatment dogs, and biopsy specimens of the left cricoarytenoideus dorsalis muscle were taken from the 4 control dogs. All biopsy specimens were blindly evaluated by histologic and histochemical evamination. At 36 to 44 weeks, the remaining 4 treatment dogs, from which biopsy specimens had not been taken were reevaluated by use of laryngoscopy with video photography. Complications and difficulties encountered during surgery included hemorrhage in the area of the cricoarytenoideus dorsalis muscle, location of a branch of the first cervical nerve that was long enough to prevent tension at the graft site, orientation of the muscle pedicle in the cricoarytenoideus dorsalis muscle without the use of an operating microscope, and preservation of the terminal portion of the first cervical nerve while forming the neuromuscular pedicle. Results of the artenoid movement evaluations, revealed improvement in arytenoid abductor function in the treatment group, compared with that in the control group at 19 weeks. Arytenoid abduction in the treatment group at this time, however, was still significantly decreased (P less than 0.05), compared with presurgical movement evaluations. Arytenoid abductor function continued to improve until it was statistically indistinguishable (P greater than 0.05) from presurgical abduction at 36 to 44 weeks. Results of the examination of biopsy specimens (neurogenic atrophy, evidence of reinnervation, or histologically normal muscle) were compatible with arytenoid movement evaluations in most of the treatment and control dogs.

OBJECTIVE To evaluate the effect of bilateral ventriculocordectomy via ventral laryngotomy on laryngeal airway resistance (LAR) in canine cadaver larynges. SAMPLE 6 clinically normal canine cadaver larynges. PROCEDURES LAR was determined for each specimen before (baseline) and after bilateral ventriculocordectomy with the epiglottis open and closed. After ventral laryngotomy was performed, the vocal cords were sharply excised, and the incised mucosal edges were apposed with 4-0 glycomer 631 suture in a simple continuous pattern. The thyroid cartilage was apposed with 3-0 polypropylene suture in a simple continuous pattern. RESULTS With the epiglottis closed, baseline median LAR was 27.6 cm H2O/L/s (range, 21.2 to 30.6 cm H2O/L/s), which did not differ significantly from the median LAR after bilateral ventriculocordectomy (24.7 cm H2O/L/s [range, 20.6 to 27.7 cm H2O/L/s]). With the epiglottis open, baseline median LAR was 7.3 cm H2O/L/s (range, 5.4 to 7.8 cm H2O/L/s), which did not differ significantly from the median LAR after bilateral ventriculocordectomy (7.2 cm H2O/L/s [range, 6.6 to 7.6 cm H2O/L/s]). CONCLUSIONS AND CLINICAL RELEVANCE Bilateral ventriculocordectomy did not affect LAR with an open epiglottis in canine cadaver larynges. Therefore, it may not be an effective treatment for laryngeal paralysis. It also did not affect LAR with a closed epiglottis, which may indicate protection against aspiration pneumonia.

OBJECTIVE To evaluate gene expression and DNA copy number in adipose tissue-derived stromal cells (ADSCs) and in ADSC-derived neurosphere-like cell clusters (ADSC-NSCs) generated from tissues of chronically paraplegic dogs. ANIMALS 14 client-owned paraplegic dogs. PROCEDURES Dorsal subcutaneous adipose tissue (< 1 cm3) was collected under general anesthesia; ADSCs were isolated and cultured. Third-passage ADSCs were cultured in neural cell induction medium to generate ADSC-NSCs. Relative gene expression of mesenchymal cell surface marker CD90 and neural progenitor marker nestin was assessed in ADSCs and ADSC-NSCs from 3 dogs by quantitative real-time PCR assay; expression of these and various neural lineage genes was evaluated for the same dogs by reverse transcription PCR assay. Percentages of cells expressing CD90, nestin, glial fibrillary acidic protein (GFAP), and tubulin β 3 class III (TUJ1) proteins were determined by flow cytometry for all dogs. The DNA copy number stability (in samples from 6 dogs) and neural cell differentiation (14 dogs) were assessed with array-comparative genomic hybridization analysis and immunocytochemical evaluation, respectively. RESULTS ADSCs and ADSC-NSCs expressed neural cell progenitor and differentiation markers; GFAP and microtubule-associated protein 2 were expressed by ADSC-NSCs but not ADSCs. Relative gene expression of CD90 and nestin was subjectively higher in ADSC-NSCs than in ADSCs. Percentages of ADSC-NSCs expressing nestin, GFAP, and TUJ1 proteins were substantially higher than those of ADSCs. Cells expressing neuronal and glial markers were generated from ADSC-NSCs and had no DNA copy number instability detectable by the methods used. CONCLUSIONS AND CLINICAL RELEVANCE Results suggested ADSCs can potentially be a safe and clinically relevant autologous source for canine neural progenitor cells. Further research is needed to verify these findings.