A Gly98Val mutation in the N-Myc downstream regulated gene 1 (NDRG1) in Alaskan Malamutes with polyneuropathy.

The first cases of early-onset progressive polyneuropathy appeared in the Alaskan Malamute population in Norway in the late 1970s. Affected dogs were of both sexes and were ambulatory paraparetic, progressing to non-ambulatory tetraparesis. On neurologic examination, affected dogs displayed predominantly laryngeal paresis, decreased postural reactions, decreased spinal reflexes and muscle atrophy. The disease was considered eradicated through breeding programmes but recently new cases have occurred in the Nordic countries and the USA. The N-myc downstream-regulated gene (NDRG1) is implicated in neuropathies with comparable symptoms or clinical signs both in humans and in Greyhound dogs. This gene was therefore considered a candidate gene for the polyneuropathy in Alaskan Malamutes. The coding sequence of the NDRG1 gene derived from one healthy and one affected Alaskan Malamute revealed a non-synonymous G>T mutation in exon 4 in the affected dog that causes a Gly98Val amino acid substitution. This substitution was categorized to be "probably damaging" to the protein function by PolyPhen2 (score: 1.000). Subsequently, 102 Alaskan Malamutes from the Nordic countries and the USA known to be either affected (n = 22), obligate carriers (n = 7) or healthy (n = 73) were genotyped for the SNP using TaqMan. All affected dogs had the T/T genotype, the obligate carriers had the G/T genotype and the healthy dogs had the G/G genotype except for 13 who had the G/T genotype. A protein alignment showed that residue 98 is conserved in mammals and also that the entire NDRG1 protein is highly conserved (94.7%) in mammals. We conclude that the G>T substitution is most likely the mutation that causes polyneuropathy in Alaskan Malamutes. Our characterization of a novel candidate causative mutation for polyneuropathy offers a new canine model that can provide further insight into pathobiology and therapy of human polyneuropathy. Furthermore, selection against this mutation can now be used to eliminate the disease in Alaskan Malamutes.

Evaluation of platelet aggregometry in dogs using the Multiplate platelet analyzer: impact of anticoagulant choice and assay duration.

OBJECTIVE: To investigate the performance of the Multiplate platelet function analyzer with regards to: (1) the use of 3 different anticoagulants (ie, citrate, hirudin, and heparin) and (2) the evaluation of optimal assay time. DESIGN: Prospective observational in vitro study. SETTING: University veterinary teaching hospital. ANIMALS: Twenty clinically healthy dogs and 3 ill dogs. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: A total of 184 analyses were performed with duplicate measurements in each test cell and results are reported as mean of the 2 measurements. Analyses were performed on blood samples from 20 dogs collected in citrate, hirudin, or heparin. A total of 4 analyses were performed on every blood sample using adenosine diphosphate, collagen, and arachidonic acid as agonists as well as a control with 0.9% sodium chloride (buffer). Aggregation in hirudin samples was significantly increased compared with heparin at all analysis times except at 6 minutes when using ADP as agonist; however, hirudin samples also demonstrated significant aggregation in the buffer control, compared to both citrate and heparin. Citrated samples yielded significantly lower aggregation compared with both hirudin- and heparin-stabilized samples at 6 and 12 minutes when ADP and collagen were used as agonists, and at most analysis times with arachidonic acid. The assay performed best at shorter analyses times, whereas longer analyses times yielded larger variation in data. CONCLUSIONS: There was a good aggregation response and acceptable analytical variation in both heparin- and hirudin-anticoagulated samples with all tested agonist at the concentrations recommended by the manufacturer. The results suggest that heparin may be superior as anticoagulant for Multiplate analyses in dogs and that short analyses times are preferable. Spontaneous platelet autoaggregation in hirudin samples warrants careful evaluation of results using this anticoagulant, especially at longer test times. The use of citrate is discouraged for Multiplate analyses in dogs due to a weak aggregation response.