Complex segregation analysis of familial amyloidosis in Oriental shorthair cats.

Amyloidosis in Siamese/Oriental cats is a lethal condition with variable age of clinical onset. There is no sex predisposition and clinical signs of disease usually become apparent by 1-7 years of age. In the terminal stages, the liver is enlarged and pale, and contains parenchymal hemorrhages. In the present study, pedigree data from 17 cats with clinical signs consistent with amyloidosis underwent genetic analysis. Necropsy and histopathological data were available for 10 of the 17 cats. Necropsy findings included pale, fragile and enlarged livers with capsular ruptures and parenchymal hemorrhages, and sanguineous effusions in the abdominal cavity. Congo red staining with birefringence confirmed systemic amyloidosis mostly involving the liver and thyroid gland. In four of the 10 cases, protein deposits were classified as amyloid A protein (AA-amyloid) by immunostaining. Pedigree data for all 17 affected cats indicated a familial trait. Animal threshold model analysis demonstrated that the heritability for amyloidosis was 0.56 ± 0.09 (standard error). Complex segregation analysis was used for statistical comparisons among models to determine environmental or sex dependent effects, and Mendelian, polygenic, or mixed Mendelian and polygenic inheritance patterns. A mixed model with a Mendelian and polygenic component provided the best fit to the data and thus was most likely. All other models of inheritance were rejected due to their insufficient ability to explain segregation of amyloidosis. In conclusion, we found evidence for a complex genetic basis for amyloidosis in Oriental shorthair cats.

Genome-wide association analysis for lethal brachycephalic-like facial dysmorphia in Labrador Retrievers.

A GWAS was performed for inborn X-linked facial dysmorphia with severe growth retardation in Labrador Retrievers. This lethal condition was mapped on the X chromosome at 17-21 Mb and supported by eight SNPs in complete LD. Dams of affected male puppies were heterozygous for the significantly associated SNPs and male affected puppies carried the associated alleles hemizygously. In the near vicinity to the associated region, RPS6KA3 was identified as a candidate gene causing facial dysmorphia in humans and mice known as Coffin-Lowry syndrome. Haplotype analysis showed significant association with the phenotypes of all 18 animals under study. This haplotype was validated through normal male progeny from a dam with the not-associated haplotype on both X chromosomes but male affected full-sibs with the associated haplotype.

[Congenital embryonal rhabdomyosarcoma of the head in a red and white German Holstein calf]. / Kongenitales embryonales Rhabdomyosarkom am Kopf eines rotbunten Deutsche-Holstein-Kalbes.

Tumours with skeletal-muscle differentiation are rare in companion animals. They are differentiated into benign rhabdomyomas and malignant rhabdomyosarcomas. A female German Holstein calf displayed a congenital, spherical, subcutaneous mass at the lateral side of the head. Histology revealed an encapsulated, expansile, highly cellular mass consisting of a reticular meshwork of moderately pleomorphic, small spindle-shaped to round cells within a fibrovascular to myxoid stroma as well as multifocal, large, blunt, multinucleated myotube-like cells (strap cells). Electron microscopy demonstrated characteristic cytoplasmic bundles of myofilaments and Z-stripes within the strap cells. Immunoreactivity for vimentin was observed in the small spindle-shaped cells and for desmin in the strap cells. The results are consistent with the spectrum of findings characteristic for a bovine congenital embryonal rhabdomyosarcoma.

Hereditary polyneuropathy in the Alaskan Malamute.

OBJECTIVE: To prove the hypothesis that a polyneuropathy in Alaskan Malamutes has a genetic background. MATERIAL AND METHODS: Pedigrees of 131 related Alaskan Malamutes were included in the current study. Neurological examination, electrodiagnosis as well as muscle and nerve biopsies could be performed in 10 dogs. Information about the disease status of the other 121 Alaskan Malamutes were supplied by referring veterinarians, breeders and owners. Segregation analysis using four different models (monogenic, polygenic, mixed monogenic-polygenic and the phenotypic model) was performed on 71 dogs to test the different mechanisms of genetic transmission. RESULTS: In seven clinically affected dogs abnormal electromyographic findings and reduced nerve conduction velocity were detected. Suspected diagnosis of polyneuropathy was confirmed by nerve biopsy results, characterized by axonal degeneration and hypomyelination. Muscle specimens revealed signs of neurogenic myopathy. Three related clinically normal Alaskan Malamutes also displayed moderate neuromuscular changes in histopathology. In the segregation analysis the polygenic model proved as best suitable to explain the observed segregation pattern among all other models tested. CONCLUSION: The current study could demonstrate that polyneuropathy in Alaskan Malamutes is a hereditary disease with variable phenotypic expression ranging from severely affected to subclinical forms, which has to be considered in future gene analysis studies.

Selection for conformation and conformational homogeneity of litters in the German shepherd dog.

Breeding standards of most dog breeds specify tolerable ranges of certain conformation traits. In the German shepherd dog (GSD), current means for withers height (WH) and BW are close to the upper breed limits. Therefore, strategies to avoid a further increase in size and to maximize the proportion of dogs fitting the breeding standard with respect to WH and BW should be compared. Body measurements were available for 14,416 male and 21,612 female GSD from 26,155 litters. Withers height and body mass index (BMI; i.e., BW/WH(2)) were considered direct selection traits. Using information on 17,154 GSD from litters with at least 2 dogs with conformation data, we defined within-litter variances of WH (vWH) and BMI (vBMI) as traits to select for the conformational homogeneity of litters. Officially recorded scores for canine hip dysplasia (CHD) of all dogs were used to monitor possible side effects of conformation selection on CHD. Genetic parameters were estimated multivariately in linear animal models by using Gibbs sampling. Heritabilities ranged between 0.19 and 0.34 for all traits, and additive genetic correlations between WH and vWH were -0.11 and those between BMI and vBMI were 0.11. The expected selection response was studied using the relative breeding values (RBV) of parents, assuming exclusion of sires, dams, or both with RBV larger than 120 and comparing means of WH, BMI, and CHD scores between offspring of all and selected parents. Concurrent selection for small WH and vWH was found to reduce the mean WH in males and females most efficiently while having little effect on CHD distributions. Multiple-trait selection for WH, vWH, BMI, and vBMI was hindered by the negative genetic correlations between the traits, and it tended to interfere with improvement of CHD status. Use of the RBV for WH and vWH for conformation selection is therefore recommended to maximize breeding success with regard to conformation and conformational homogeneity in the GSD.

Genetic analyses of elbow and hip dysplasia in the German shepherd dog.

Results from radiographic screening for canine hip dysplasia (CHD) and elbow dysplasia (CED) of 48 367 German shepherd dogs born in 2001-07 were used for the population genetic analyses. Available information included CHD scores for 47 730 dogs, CED scores for 28 011 dogs and detailed veterinary diagnoses of primary ED lesions for a subsample of 18 899 dogs. Quasi-continuous traits were CHD, CED and cases of CED without radiographically visible primary lesion (CED-ARTH). Binary coding was used for fragmented medial coronoid process of the ulna (FCP), borderline findings and mild to severe signs of dysplasia in hip and elbow joints. Genetic parameters were estimated in univariate threshold and multivariate linear and mixed linear-threshold models using Gibbs sampling. Correlations between univariately predicted breeding values (BV) indicated genetic differences between borderline and affected disease status for both CHD (r(BV) = 0.5) and CED (r(BV) = 0.3). Multivariate genetic analyses with separate consideration of borderline findings revealed moderate heritabilities of 0.2-0.3 for the quasi-continuous traits with positive additive genetic correlation of 0.3 between CHD and both CED and CED-ARTH. For FCP, heritability of 0.6 and additive genetic correlations of +0.1 to CHD and -0.1 to CED-ARTH were estimated. Results supported the relevant genetic determination of CHD and CED, argued for both diseases against interpretation of borderline findings as healthy and implied genetic heterogeneity of CED. Accordingly, future breeding strategies to reduce the prevalences of CHD and CED in the German shepherd dog should be most efficient when based on BV from multivariate genetic evaluation for CHD, CED-ARTH and FCP with use of the whole scale of categories for classification of CHD and CED.

Simulation study on the effects of excluding offspring information for genetic evaluation versus using genomic markers for selection in dog breeding.

Different modes of selection in dogs were studied with a special focus on the availability of disease information. Canine hip dysplasia (CHD) in the German shepherd dog was used as an example. The study was performed using a simulation model, comparing cases when selection was based on phenotype, true or predicted breeding value, or genomic breeding value. The parameters in the simulation model were drawn from the real population data. The data on all parents and 40% of their progeny were assumed to be available for the genetic evaluation carried out by Gibbs sampling. With respect to the use of disease records on progeny, three scenarios were considered: random exclusion of disease data (no restrictions, N), general exclusion of disease data (G) and exclusion of disease data for popular sires (P). One round of selection was considered, and the response was expressed as change of mean CHD score, proportion of dogs scored as normal, proportion of dogs scored as clearly affected and true mean breeding value in progeny of popular sires in comparison with all sires. When no restrictions on data were applied, selection on breeding value was three times more efficient than when some systematic exclusion was practised. Higher selection response than in the exclusion cases was achieved by selecting on the basis of genomic breeding value and CHD score. Genomic selection would therefore be the method of choice in the future.

Effect of polymorphisms in four candidate genes for fertility on litter size in a German pig line.

We carried out an SNP discovery project in pigs for candidate genes playing potentially important roles in embryonic development. Using eight pigs one each from eight breeds (Meishan, Mangalitza, Duroc, Pietrain, German Landrace, Hampshire, Husum Red Pied, German Large White), 36 SNPs were identified in intronic sequences of 21 porcine candidate genes based on sequencing of PCR products. The primer pairs were designed using porcine EST sequences allowing amplification of introns. These SNPs were tested for their association with the number of piglets born alive in German Large White sows using a discordant approach. Significant effects (p < 0.001 and p < 0.05, respectively) of intronic SNPs on litter size were found for four genes: mitogen-activated protein kinase kinase kinase 3 (MAP3K3), vascular endothelial growth factor receptor (KDR), erbb2 interacting protein (ERBB2IP) and peroxisome proliferator-activated receptor delta (PPARD). These SNPs can be further tested in upcoming association studies for their influence on litter size in different breeds using larger sample sizes.

INHBA-associated markers as candidates for stallion fertility.

The inhibin beta A (INHBA) gene was chosen as candidate for stallion fertility and analysed for intragenic markers to find associations with pregnancy rate per oestrus. Intragenic single nucleotide polymorphisms (SNPs) were developed in order to perform an association and haplotype analysis using the least square means (LSM) of the pregnancy rate per oestrus for stallions as well as breeding values (BVs) for the embryonic and paternal component of the pregnancy rate per oestrus. The polymorphisms were genotyped in 161 Hanoverian warmblood stallions. Insemination records from approximately 20,000 Hanoverian warmblood mares were used to calculate LSM for stallions and to predict the paternal and embryonic component of BVs for the pregnancy rate per oestrus. We demonstrated significant associations of single markers and haplotypes with the LSM and the embryonic and paternal component of BVs for the pregnancy rate per oestrus. This is the first report on INHBA as an associated candidate gene with the LSM of stallions and the paternal and embryonic component of BVs for the pregnancy rate per oestrus.

Effect of polymorphisms in the genes for LIF and RBP4 on litter size in two German pig lines.

The association of two diallelic polymorphisms in the porcine genes for leukaemia inhibitory factor (LIF) and retinol-binding protein 4 (RBP4) with number of piglets born alive (NBA) in two German pig lines was studied. The investigated single nucleotide polymorphism (SNP) in the porcine LIF gene has been located in the 3'-untranslated region of its third exon, whereas the SNP in the RBP4 gene genotyped in this study is intronic. At the LIF locus the allele frequencies were 0.613 for the A allele and 0.387 for the B allele in German Landrace (GL) and 0.276 for A and 0.724 for B in German Large White (GW). At the RBP4 locus, the allele frequencies were 0.586 for the A allele and 0.414 for the B allele in GL and 0.733 for A and 0.267 for B in GW. There was a significant additive effect of the LIF B allele on NBA in GW over all parities (p

Review on canine elbow dysplasia: pathogenesis, diagnosis, prevalence and genetic aspects.

Elbow dysplasia (ED) is a progressive skeletal disease, which may comprise osteoarthrotic changes, incongruity of the joint, a fragmented medial coronoid process (FCP), osteochondrosis dissecans of the trochlea humeri (OCD), and an ununited anconeal process (UAP). Disturbances of enchondral ossification, as well as asynchronous growth of the antebrachial bones and an increased growth rate may provoke abnormal stresses on specific locations such as the medial coronoid process, the anconeal process of the ulna or the humeral trochlea, which may result in ED. Overnutrition with calcium, vitamin D and energy had important influence on rapidity of growth and proneness to ED. ED manifests most often in young dogs less than 1 1/2 year, causing clinical signs such as (intermittent) lameness, pain on movement and altered posture of the affected limb, and radiographically secondary osteoarthrotic changes such as osteophytes or the aforementioned primary lesions. As radiographic projection of the primary lesions FCP and OCD may be difficult in the commonly used mediolateral flexed and craniocaudal views, additional projections such as mediolateral extended or anteroposterior oblique views and alternative diagnostic means such as computed tomography (CT) may be worth considering. Cases of ED are reported in various breeds, though some breeds are especially predisposed and seem to be prone to specific primary lesions. For the German shepherd dog, a breed-predisposition for UAP may be explained by broad chondral junction in association with an accelerated pattern of skeletal maturation. Heritabilities were estimated in a variety of studies, depending strongly on the type of model used. The mode of inheritance is not yet ascertained, but recently the common assumption of a polygenic mode of inheritance for ED is doubted. Instead, genetic independence was supposed between different primary lesions, and also for ED as well as for FCP major gene influence was discussed. So long, phenotypic mass selection was accomplished in different countries most often resulting in a reduced prevalence of ED. The use of best linear unbiased prediction (BLUP) methods including information on relatives and separate evaluation of genetically independent primary lesions may further improve selection against ED.

A polymorphism within the equine CRISP3 gene is associated with stallion fertility in Hanoverian warmblood horses.

Fertility of stallions is of high economic importance, especially for large breeding organisations and studs. Breeding schemes with respect to fertility traits and selection of stallions at an early stage may be improved by including molecular genetic markers associated with traits. The genes coding for equine cysteine-rich secretory proteins (CRISPs) are promising candidate genes because previous studies have shown that CRISPs play a role in the fertilising ability of male animals. We have previously characterised the three equine CRISP genes and identified a non-synonymous polymorphism in the CRISP1 gene. In this study, we report one non-synonymous polymorphism in the CRISP2 gene and four non-synonymous polymorphisms in the CRISP3 gene. All six CRISP polymorphisms were genotyped in 107 Hanoverian breeding stallions. Insemination records of stallions were used to analyse the association between CRISP polymorphisms and fertility traits. Three statistical models were used to evaluate the influence of single mutations, genotypes and haplotypes of the polymorphisms. The CRISP3 AJ459965:c.+622G>A SNP leading to the amino acid substitution E208K was significantly associated with the fertility of stallions. Stallions heterozygous for the CRISP3 c.+622G>A SNP had lower fertility than homozygous stallions (P = 0.0234). The pregnancy rate per cycle in these stallions was estimated to be approximately 7% lower than in stallions homozygous at this position.

A high-resolution comparative radiation hybrid map of equine chromosome 4q12-q22.

In this study, we present a comprehensive 5000-rad radiation hybrid map of a 40-cM region on equine chromosome 4 (ECA4) that contains quantitative trait loci for equine osteochondrosis. We mapped 29 gene-associated sequence tagged site markers using primers designed from equine expressed sequence tags or BAC clones in the ECA4q12-q22 region. Three blocks of conserved synteny, showing two chromosomal breakpoints, were identified in the segment of ECA4q12-q22. Markers from other segments of HSA7q mapped to ECA13p and ECA4p, and a region of HSA7p was homologous to ECA13p. Therefore, we have improved the resolution of the human-equine comparative map, which allows the identification of candidate genes underlying traits of interest.

Inheritance of canine hip dysplasia: review of estimation methods and of heritability estimates and prospects on further developments.

Canine hip dysplasia is a long-known, widespread degenerative skeletal disease. A hereditary component of hip dysplasia was assumed early, although attempts to explain hip dysplasia with known Mendelian modes of inheritance did not sufficiently fit the data observed. Nevertheless, both recessive and dominant modes of inheritance were proposed. Later on, it was proposed that CHD was determined in a multifactorial way. Both the influence of many genes and environmental effects were assumed to affect the development of CHD. More recently, this thesis was supplemented and refined, as a major gene was detected as a cause of CHD in addition to a polygenic component. Nowadays, projects are under way with the aim to locate quantitative trait loci (QTL) significantly linked to CHD, and ultimately to develop gene tests to identify carriers of genes responsible for CHD.

Genetic analysis of three different classification protocols for the evaluation of elbow dysplasia in German shepherd dogs.

OBJECTIVES: Three different scoring systems for elbow dysplasia and its radiographic signs were genetically evaluated in 2645 German shepherd dogs. METHODS: An animal model was used to estimate heritabilities and additive genetic and residual correlations for the three scoring systems: ED-SV, which is recommended by the International Elbow Working Group; ED-LA, developed by Lang and others; and ED-TH, proposed by Tellhelm. RESULTS: The effects of sex, age at examination and the correlation between the two factors were significant for all three scoring systems. Heritability estimates (se) were 0.18 (0.04) for ED-SV, 0.11 (0.03) for ED-LA and 0.16 (0.04) for ED-TH. The additive genetic correlations among the different single criteria for elbow dysplasia and the different elbow dysplasia scores were between 0.68 and 0.98, except for the criteria ununited anconeal process and osteochondrosis dissecans of the trochlea humeri, which were mostly genetically negatively correlated to the other radiological criteria. CLINICAL SIGNIFICANCE: The elbow dysplasia scores were determined by two genetically different traits. The possibilities for selecting German shepherd dogs with respect to elbow dysplasia might be improved by taking into account these two traits in the prediction of breeding values.

Complex segregation analysis of canine hip dysplasia in German shepherd dogs.

Complex segregation analyses were carried out to clarify the mode of inheritance of canine hip dysplasia (CHD) in German shepherd dogs. Data were used from 8,567 animals examined for CHD from 20 families with three to four generations. The existence of a major gene in addition to polygenic gene effects was detected. In the present study, a mixed model with a dominant major gene effect seemed to be most probable for dichotomous encoding (0: dogs without signs of CHD; 1: dogs with borderline/slight to severe CHD). In addition, mixed major gene inheritance was shown for a binary trait where borderline was assigned to dogs scored free from CHD and for a trichotomously encoded trait (0: dogs without signs of CHD; 1: borderline CHD; 2: mild to severe CHD). Although only small frequencies were found for the unfavorable homozygotic genotype AA, the probability of the AB genotype was high in affected animals. Selection schemes to reduce the frequency of the allele A should therefore efficiently improve existing breeding programmes in German shepherd dogs.