Deafness in Australian Cattle Dogs associated to QTL on chromosome 20 in genome-wide association study analyses.

Pigment-associated deafness is a common hereditary condition in a range of dog breeds. The aim of this study was to perform a genome-wide association analysis to investigate the genetic architecture of deafness in Australian Cattle Dogs. Genotypes for 104 757 polymorphisms in 216 dogs were available for analyses after quality control. A genomic relationship matrix was used in the mixed model analyses to account for polygenic effects, as we tested each polymorphism for its association with deafness, in a case/control experimental design. Three approaches were used to code the genotypes and test for additive, recessive and dominant SNP effects. The genome-wide association study analyses identified a clear association peak on CFA20, with the most significant SNPs on this chromosome (1.29 × 10-4 ) in the vicinity of MITF. Variants in MITF have been associated with white pigmentation in dogs and with deafness in humans and other species, supporting the premise that canine deafness is associated with variants in or near this gene. A recessive inheritance for the peak in CFA20 is possible given the significant results in the recessive model; however, the estimated heritability was low (4.54 × 10-5 ). Further validation, identification of variants and testing in other dog breeds are needed.

Prevalence of deafness in dogs heterozygous or homozygous for the merle allele.

BACKGROUND: Deafness in dogs is frequently associated with the pigment genes piebald and merle. Little is known about the prevalence of deafness in dogs carrying the merle allele. OBJECTIVE: To determine the prevalence of deafness in dogs heterozygous and homozygous for the merle allele of the mouse Silver pigment locus homolog (SILV) gene. ANIMALS: One hundred and fifty-three privately owned merle dogs of different breeds and both sexes. METHODS: Hearing was tested by brainstem auditory-evoked response and classified as bilaterally hearing, unilaterally deaf, or bilaterally deaf. DNA from buccal cells was genotyped as either heterozygous or homozygous for the merle allele. Deafness association tests among merle genotype, eye color, and sex were performed by the chi(2) test. RESULTS: Deafness prevalence in merles overall was 4.6% unilaterally deaf and 4.6% bilaterally deaf. There was a significant association between hearing status and heterozygous versus homozygous merle genotype. For single merles (Mm), 2.7% were unilaterally deaf and 0.9% were bilaterally deaf. For double merles (MM), 10% were unilaterally deaf and 15% were bilaterally deaf. There was no significant association with eye color or sex. CONCLUSIONS: Deafness prevalence in merle dogs was greater than that in some dog breeds homozygous for the piebald gene, such as the English Cocker Spaniel, but comparable to, or lower than, that in the Dalmatian and white Bull Terrier. Dogs homozygous for the merle allele were significantly more likely to be deaf than heterozygotes.

Distortion product otoacoustic emissions in geriatric dogs.

Recordings of distortion product otoacoustic emissions (DPOAE) were taken from 28 geriatric dogs aged 12.2 ± 2.2 years and 15 control dogs aged 5.9 ± 3.0 years (mean ± standard deviation) to demonstrate frequency-specific changes in cochlear responses. Recordings were performed for primary frequencies of 2-12 kHz in 2 kHz increments. Brainstem auditory evoked response (BAER) recordings were also made from geriatric dogs for comparison with DPOAE responses. Significant decreases in DPOAE response amplitudes were observed at frequencies of 6-12 kHz in geriatric dogs compared to control dogs, reflecting loss of cochlear outer hair cells along the length of the cochlea. Significant decreases in response amplitudes were not seen at frequencies of 2 or 4 kHz. Decreases in BAER response amplitudes subjectively paralleled the depressed DPOAE amplitudes. No significant linear regression relationships were found for DPOAE response amplitude vs. age despite the progressive nature of age-related hearing loss. The reductions in response at all frequencies starting at the age where dogs are considered geriatric indicate that age-related hearing loss begins earlier in the life span. DPOAE recordings provide a means to assess cochlear function across different portions of the auditory spectrum for assessing hearing loss associated with aging, and potentially for losses from other causes of decreased auditory function.

Heritability and segregation analysis of deafness in U.S. Dalmatians.

Hereditary loss of hearing affects many breeds of the domestic dog, but the Dalmatian has the highest prevalence. Approximately 30% are affected in the United States (U.S.) population. It is widely accepted that a relationship exists between deafness and pigmentation in the dog and also in other animals. While the Dalmatian exemplifies this relationship, the genetic origin and mode of inheritance of deafness in this breed are unknown. The goals of this study were to: (1) estimate the heritability of deafness in an extended kindred of U.S. Dalmatians and (2) determine, through complex segregation analysis, whether there is a major segregating locus that has a large effect on the expression of deafness. A kindred of 266 Dalmatians was assembled, of which 199 had been diagnosed using the brainstem auditory evoked response to determine auditory status. Of these, 74.4% (N = 148) had normal hearing, 18.1% (N = 36) were unilaterally deaf, and 7.5% (N = 15) were bilaterally deaf. A heritability of 0.73 was estimated considering deafness a dichotomous trait and 0.75 considering it as a trichotomous trait. Although deafness in the Dalmatian is clearly heritable, the evidence for the presence of a single major gene affecting the disorder is not persuasive.

Inhibition of synaptosomal uptake of amino acid transmitters by diamines.

Reports of the inhibitory effects of diaminocarboxylic acids on the uptake of amino acid transmitters led the present authors to examine the effects of simple aliphatic diamines on the synaptosomal uptake of glutamate, aspartate, GABA and glycine. The diamines studied were the series from ethylenediamine through to 1,7-diaminoheptane; DL-2,4-diaminobutyric acid (DABA) was also tested for comparative purposes. The greatest inhibition seen was on the uptake of glycine and GABA. Weaker effects on uptake were seen with glutamate, while aspartate was unaffected. The patterns of inhibition for glycine and GABA were similar and the effects were dose-dependent. 1,2-Diaminopropane was the most inhibitory, followed by ethylenediamine and 1,7-diaminoheptane. The reported inhibitory effects of DL-2,4-diaminobutyric acid on the uptake of GABA and glutamate were confirmed; comparable inhibition of the uptake of glycine and aspartate was seen but the effects on GABA were most potent. Inhibition of the uptake of GABA by 1,2-diaminopropane was approximately one fifteenth that reported for DL-2,4-diaminobutyric acid. The inhibition by diamine of the uptake of glycine and GABA can provide an explanation of the depressant effects of diamines, seen after ventricular administration; however, the excitotoxic effects of the diamines 1,3-diaminopropane through to 1,7-diaminoheptane could not be explained by the present results.

N-CAM promotes recovery in injured nerves.

The neural cell adhesion molecule, N-CAM, makes critical contributions to the development of the nervous system. It mediates the stability of homophilic adhesion in embryonic neurons and participates in morphologic differentiation. The goal of these studies was to determine N-CAM contributions to nerve regeneration and recovery of function in two species with an excised segment of sciatic nerve. N-CAM was isolated from embryonic brains, affinity purified and admixed in collagen gel for administration. Recovery was compared 30 days after surgery for two types of N-CAM delivery: entubulization versus direct application. For control nerves, tubes contained gel only. In preliminary chicken studies, latency of nerve responses was measured to demonstrate N-CAM's ability to improve upon spontaneous recovery. In subsequent studies of rodent nerves, the direct application of N-CAM significantly improved recovery in evoked nerve response amplitude, number of regenerated axons and behavioral activity. Results demonstrate N-CAM's ability to augment nerve regeneration and suggest a potential for therapeutic use.

Flash and pattern reversal visual evoked potentials in C57BL/6J and B6CBAF1/J mice.

Visual system responses (visual evoked potentials) to flash (FVEP) and pattern reversal (PRVEP) stimuli were recorded in mice. Two strains were used: black C57BL/6J mice and agouti B6CBAF1/J mice (first generation offspring of C57BL/6J females and CBA/J males.) Subjects were sedated with ketamine and xylazine. Flash rate (FVEP) and stimulus spatial frequency and pattern reversal rate (PRVEP) were varied to determine optimum stimulus parameters. Normative FVEP and PRVEP data were collected from mice of both strains after determination of optimum parameters. Five positive and four negative alternating peaks were routinely observed in the FVEP, while three positive and three negative alternating peaks were seen with the PRVEP. Varying the flash rate, the pattern reversal rate, and spatial frequency significantly affected nearly all amplitude and latency measures in the responses. Significant differences between strains were seen on some, but not all, latency and amplitude measures when the stimulus parameters were varied.

Hyperbaric oxygen effects on brown recluse spider (Loxosceles reclusa) envenomation in rabbits.

Human loxoscelism was modeled in albino rabbits by injection of brown recluse spider (Loxosceles reclusa) venom, and the effects of daily or twice-daily hyperbaric oxygen treatment on wound healing were investigated. Lesions similar to those seen in humans were produced in rabbits by intradermal injection of 200 microliters of a venom extract (0.21 microgram protein per microliter), including edema and erythema, ischemia and cyanosis in the first 12 hr, extensive purpura by 24 hr, and crateriform ulcer formation by day four, with induration and eschar formation. Hyperbaric oxygen treatments, consisting of two atmospheres absolute (2 ATA) for 60 min, were applied daily (n = 8) or twice daily (n = 8), while control animals (n = 8) received no treatment. Treatments were initiated 72 hr after venom injection (day 3) to duplicate typical clinical treatment delays, and were administered for seven consecutive days. No significant effects of hyperbaric oxygen treatment on lesion healing were seen as measured by lesion area. However, histologic evaluation of wound tissue collected at euthanasia on day 24 showed clear differences between rabbits receiving twice-daily treatments and those receiving daily or no treatment. The former showed complete re-epithelization or slight ulceration, while the latter usually had necrotic cavities extending into the dermis, with myonecrosis and inflammatory cell accumulation. Thus, no superficial differences were seen between groups, but twice-daily treatments resulted in enhanced recovery at the histologic level.

Antagonism of kainic acid lesions in the mouse hippocampus by U-54494A and U-50488H.

A morphometric study of kainic acid- (KA) induced lesions was designed for the study of the interaction of the diamines U-5449A and U-50488H with excitatory amino acids, and the dose-response relationship thereof. IC50S determined for binding at the kappa receptor and other opioid receptors demonstrated the lack of kappa activity of U-54494A, a structurally related analog of U-50488H. Both opiate kappa receptor related anticonvulsant diamines were tested for their ability to protect the mouse hippocampus from the cytopathological changes induced by KA in neurons and glia. The damage observed with i.c.v. KA in mouse was restricted to neurons of the CA3 pyramidal region and glia of the hippocampus. It involved massive cell loss and shrunken neurons with dark cytoplasm and nuclei. Groups treated with combinations of KA and U-54494A or U-50488H showed scarce damage, but patches of necrotic changes were still observed. Control animals treated with saline (i.c.v.) and U-54494A (s.c.) or U-50488H (s.c.) did not suffer any noticeable alterations of the polymorphic layers of the hippocampal formation. Image analysis of the CA3 area of the hippocampus was used to quantitate the vacuolization induced by KA lesions in the control and treated groups. By this method, both U-54494A and U-50488H were shown to protect this area in a dose-related fashion as evidenced by reduced vacuolization. The anticonvulsant properties of these compounds may result in the antagonism of the excitotoxic lesions. More specifically, the ability of these diamines to block depolarization-induced influxes of Ca++ may protect the CA3 cells from the cytotoxic effects of persistent depolarization.

Autonomic regulation of gastric ulcerogenesis after cervical cord transection in the rat.

OBJECTIVE: To examine the role of anticholinergic and sympathomimetic drugs in preventing gastric ulcerogenesis after cervical cord transection (CCT) in the rat. METHODS: A randomized, prospective, interventional trial was performed comparing pirenzepine (muscarinic type I receptor antagonist) and ephedrine (nonspecific sympathomimetic) in the prevention of gastric ulcerogenesis after CCT in the rat. After isoflurane-induced general anesthesia, group 1 (n = 12) received sham CCT with no pretreatment, group 2 received CCT with no pretreatment, group 3 received CCT with pirenzepine pretreatment (0.01/mg/kg IP), and group 4 received CCT with ephedrine pretreatment (3 mg/kg IP). Six hours after intervention, all the rats were euthanized with isoflurane, stomachs were dissected, and a gastric ulcer index was determined. RESULTS: The mean (+/- SD) ulcer index was 0.08 +/- 0.1 for group 1, 2.33 +/- 0.5 for group 2 (p = 0.01), 0.41 +/- 0.7 for group 3 (p = 0.037 compared with group 2), and 0.75 +/- 0.7 for group 4 (p = 0.0005 compared with group 2). Groups 3 and 4 were not significantly different from each other (p = 0.30). CONCLUSIONS: Gastric ulcerogenesis after CCT in the rat is decreased by anticholinergic and sympathomimetic drug pretreatment.

Visual evoked potentials in the clinically normal dog.

Visual evoked potentials (VEP) in response to flashes of white light were recorded from 15 adult beagles of both sexes to provide a normative data base. Separate recordings were taken by stimulating each eye of every dog. Responses were recorded from a needle electrode placed over the nuchal crest referenced to an electrode just caudal to the eyes. Five positive and negative peaks were present in each VEP; P1, N1, P2, N2, and P3. Peak P2 was the most prominent. Mean (+/- standard deviation [SD]) latencies for peaks P1, N1, P2, N2, and P3 were 14.3 +/- 2.4, 29.2 +/- 2.2, 54.5 +/- 7.4, 78.0 +/- 13.1, and 98.1 +/- 12.6 msec, respectively. Peak-to-peak mean amplitudes ranged from 5.88 to 13.30 microV. Recordings were accomplished without sedation, anesthesia, or mydriatic drugs.

Postnatal development of brainstem auditory-evoked potentials, electroretinograms, and visual-evoked potentials in the calf.

Brainstem auditory-evoked potentials (BAEP), electroretinograms (ERG), and visual-evoked potentials (VEP) were recorded for eight calves from birth to 56 days and the values compared with previously determined adult responses. The BAEPs, ERGs, and VEPs recorded within the first 24 hours after birth contained all of the peaks seen in adult recordings. Varying degrees of maturation of the responses were documented as changes in latency and amplitude with age. The BAEPs were adult-like at birth, with latencies falling within the mean, plus or minus one standard deviation, for adult cows. A small but significant decrease in latency with age was seen for the first, second, and fourth peaks of the response. The ERG amplitudes were also within the adult range for the entire period of the study. Latencies to the a- and b-waves declined during the first 14 days and then stabilized at adult values. The VEP latencies decreased with age, with late peaks changing more than early peaks. Latencies of all but the first peak decreased to values less than the adult range. Two VEP amplitudes increased significantly with age. Developmental appears in the calf and other precocious species are compared to those in altricious (nonprecocious) species.

Brainstem auditory-evoked potentials in Holstein cows.

Brainstem auditory-evoked potentials (BAEP) were recorded from 29 Holstein cows in a typical clinical setting. The latencies of five positive peaks in the BAEP were measured, and latency-intensity functions were determined. The BAEP was similar to that reported in humans, dogs, horses, and other species. The responses were reproducible for each cow, with low variability between cows. Four peaks (I, II, III, V) were present in all recordings, and a fifth (IV) was present infrequently. All peak latencies decreased as click-stimulus intensity increased. The threshold for detection of the BAEP was higher than expected for the cow compared with the horse.

Brainstem auditory-evoked potential assessment of congenital deafness in Dalmatians: associations with phenotypic markers.

To screen for congenital deafness, brainstem auditory-evoked potential (BAEP) testing was performed on 1031 Dalmatians from three geographically separated areas. Phenotypic marker assessment was done to determine markers possibly associated with deafness. Markers included sex, hair coat color, pigmentation of different areas of skin (eye rims, nose, and ears), presence of a patch, spot size and marking (density of spotting), sire and dam BAEP status, and presence of iris and retinal tapetal pigmentation. Combined data from all test sites showed 8.1% bilateral deafness (N = 83 dogs) and 21.6% unilateral deafness (N = 223), or an overall 29.7% incidence of hearing disorders. Significant (P less than 0.05) associations with deafness for the data from all test sites combined were seen for patch, sire and dam BAEP, iris pigment, and retinal pigment. However, results differed for several of the significant phenotypic markers when analyses were done on the data from the individual test sites; changes from significant to not significant were found. This suggested the existence of multiple populations of deafness patterns, and reinforced the precautionary conclusion that associations of phenotypic markers with deafness are not necessarily functionally significant.

Electroretinogram and visual-evoked potential measurements in sheep.

Electroretinogram (ERG) and visual-evoked potential (VEP) recordings were taken from ten Suffolk-cross sheep. Stimuli for VEP were 1.5 flashes of white light/s; ERG stimuli were single flashes. The ERG measurements of the a and b wave latencies and a-to-b amplitude were measured between the lower eyelid and the vertex, with ground on the nuchal crest. The VEP after monocular stimulation were measured between the nuchal crest and the interorbital line, with ground on the vertex. Measurements consisted of the latencies to seven alternating positive and negative peaks P1, N1, P2, N2, P3, N3 and P4, and six amplitudes, P1-N1, N1-P2, P2-N2, N2-P3, P3-N3 and N3-P4. Average latencies for the a and b waves were 13.6 and 28.2 ms; the mean ab amplitude was 131.68 microV. Average latencies for the seven VEP peaks were 35.0, 43.1, 52.8, 64.1, 74.5, 90.4 and 112.2 ms. Mean amplitudes ranged from 3.90 to 8.29 microV.

Toxic Bermuda grass tremor in the goat: an electroencephalographic study.

Tremorgenic Bermuda grass hay harvested during the 1971 Louisiana outbreak of toxicosis and kept in cold storage until 1979 was fed to a female Nubian cross goat. Cortical EEG from chronically implanted screw electrodes was recorded daily during hay consumption. Typical clinical signs of toxicosis were manifested as ataxia on day 8 and tremor on day 11. Motor activity returned to normal 2 days after reinstitution of nontoxic Bermuda grass hay. Changes in EEG amplitudes, frequency content, or wave-forms were not seen during the development or after the appearance of toxicosis. The administration of diazepam at the height of toxicosis suppressed the tremor for several hours.

Serum and cerebrospinal fluid concentrations of immunoglobulin G in Suffolk sheep with scrapie.

Determinations were made by laser nephelometry of serum and CSF immunoglobulin (Ig) G concentrations in Suffolk sheep with naturally occurring scrapie. The serum IgG concentrations in 3 sheep with confirmed or suspected scrapie were between 2,140 and 3,290 mg of IgG/100 ml, and the CSF values were between less than 10 and 75 mg of IgG/100 ml. In 8 clinically healthy (control) sheep, serum IgG concentrations were 2,647 to 7,380 mg/100 ml and CSF IgG concentrations were between 0 (undetectable) and 162 mg/100 ml. A sheep with pulmonary adenomatosis had 1,445 mg of IgG/100 ml of serum. The results indicated that neither serum nor CSF IgG concentrations were increased in sheep with naturally occurring infection with scrapie and that the severity of the disease did not correspond with the IgG concentration.

Postnatal development of the brain stem auditory-evoked potential in dogs.

Recordings of averaged brain stem auditory-evoked potentials were obtained from 13 Beagle pups of both genders to document the postnatal development of the response from age 1 to 76 days. Responses were recorded between needle electrodes placed on the vertex and the ipsilateral ear, with ground at the interorbital line. Recordings were performed without sedation. Low-amplitude responses to high-intensity stimuli could be recorded from animals prior to opening of the ear canals. Peak latencies did not change after day 20 for peak I, day 30 for peaks II and III, and day 40 for peak V. As a result, the interpeak latencies between peaks I and III did not change after day 30, but continued to decrease until day 40 for peaks III-V and I-V. Peak amplitudes reached plateau values by day 20 (peak I) or day 30 (peaks II, III, and V). All of the measured latency and amplitude values had significant (P less than 0.001) linear regression lines of latency vs age and amplitude vs age. The brain stem auditory-evoked potential thresholds were mature by day 20.

Electroretinogram and visual-evoked potential measurements in Holstein cows.

The electroretinogram (ERG) and flash visual-evoked potential (FVEP) were recorded from 25 Holstein cows in a clinical setting without dark adaptation. Latencies to the a and b waves of the ERG were measured bilaterally. The ERG recordings were between the lower eyelid of the stimulated eye and the vertex, with ground on the nuchal crest. Stimuli for ERG and FVEP were 1.5 flashes of white light/s. Recorded ERG were highly consistent and repeatable. The average latencies for the a and b waves were 14 and 30 ms; the mean ab amplitude was 43 microV. The FVEP were recorded bilaterally between the middle of the interocular line and the midpoint of the nuchal crest, with ground on the vertex. The FVEP measurements included latencies to 5 alternating positive and negative peaks (P1, N1, P2, N2, and P3) and 4 peak-to-peak amplitudes (P1 to N1, N1 to P2, P2 to N2, and N2 to P3). The P2 peak was consistently the most prominent. Average latencies for the 5 peaks were 46, 64, 86, 106, and 137 ms for P1, N1, P2, N2, and P3, respectively. The FVEP peak-to-peak amplitudes had a high variability.

Electroencephalogram and evoked potentials in naturally occurring scrapie in sheep.

Electroencephalogram (EEG), brain stem auditory-evoked potential, and flash visual-evoked potential recordings were taken from healthy sheep and from 3 sheep with scrapie, a CNS slow virus infection. The EEG changes included semi-periodic, polyphasic, high-voltage sharp waves (bilaterally synchronous and symmetric in all channels), and a cyclic alternating pattern consisting of a high-voltage low-frequency phase, followed by a low-voltage high-frequency phase. The high-voltage phase occurred with increased arousal, and the low-voltage phase occurred with decreased arousal. Myoclonic jerks were coincident with EEG sharp waves in one sheep with scrapie. Several spontaneous focal seizures were observed. Wave-form amplitudes were greatly reduced in the brain stem auditory-evoked potential and flash visual-evoked potential; degree of reduction did not always correlate with disease severity. The EEG and evoked potential changes were seen in an exposed sheep that had not yet developed clinical signs of scrapie.