A missense variant in the coil1A domain of the keratin 25 gene is associated with the dominant curly hair coat trait (Crd) in horse

Curly horses present a variety of curl phenotypes that are associated with various degrees of curliness of coat, mane, tail and ear hairs. Their origin is still a matter of debate and several genetic hypotheses have been formulated to explain the diversity in phenotype, including the combination of autosomal dominant and recessive alleles. Our purpose was to map the autosomal dominant curly hair locus and identify the causal variant using genome-wide association study (GWAS) and whole-genome sequencing approaches. A GWAS was performed using a Bayesian sparse linear mixed model, based on 51 curly and 19 straighthaired French and North American horses from 13 paternal families genotyped on the Illumina EquineSNP50 Bead-Chip. A single strong signal was observed on equine chromosome 11, in a region that encompasses the type I keratin gene cluster. This region was refined by haplotype analysis to a segment including 36 genes, among which are 10 keratin genes (KRT-10, -12, -20, -23, -24, -25, -26, -27, -28, -222). To comprehensively identify candidate causal variants within all these genes, whole-genome sequences were obtained for one heterozygous curly stallion and its straighthaired son. Among the four non-synonymous candidate variants identified and validated in the curly region, only variant g. 21891160G> A in the KRT25 gene (KRT25:p.R89H) was in perfect agreement with haplotype status in the whole pedigree. Genetic association was then confirmed by genotyping a larger population consisting of 353 horses. However, five discordant curly horses were observed, which carried neither the variant nor the main haplotype associated with curliness. Sequencing of KRT25 for two discordant horses did not identify any other deleterious variant, which suggests locus rather than allelic heterogeneity for the curly phenotype. We identified the KRT25: p. R89H variant as responsible for the dominant curly trait, but a second dominant locus may also be involved in the shape of hairs within North American Curly horses

Omics technologies provide new insights into the molecular physiopathology of equine osteochondrosis

Background: Osteochondrosis (OC(D)) is a juvenile osteo-articular disorder affecting several mammalian species. In horses, OC(D) is considered as a multifactorial disease and has been described as a focal disruption of endochondral ossification leading to the development of osteoarticular lesions. Nevertheless, OC(D) physiopathology is poorly understood. Affected horses may present joint swelling, stiffness and lameness. Thus, OC(D) is a major concern for the equine industry. Our study was designed as an integrative approach using omics technologies for the identification of constitutive defects in epiphyseal cartilage and/or subchondral bone associated with the development of primary lesions to further understand OC(D) pathology. This study compared samples from non-affected joints (hence lesion-free) from OC(D)-affected foals (n = 5, considered predisposed samples) with samples from OC-free foals (n = 5) considered as control samples. Consequently, results are not confounded by changes associated with the evolution of the lesion, but focus on altered constitutive molecular mechanisms. Comparative proteomics and micro computed tomography analyses were performed on predisposed and OC-free bone and cartilage samples. Metabolomics was also performed on synovial fluid from OC-free, OC(D)-affected and predisposed joints. Results: Two lesion subtypes were identified: OCD (lesion with fragment) and OC (osteochondral defects). Modulated proteins were identified using omics technologies (2-DE proteomics) in cartilage and bone from affected foals compare to OC-free foals. These were associated with cellular processes including cell cycle, energy production, cell signaling and adhesion as well as tissue-specific processes such as chondrocyte maturation, extracellular matrix and mineral metabolism. Of these, five had already been identified in synovial fluid of OC-affected foals: ACTG1 (actin, gamma 1), albumin, haptoglobin, FBG (fibrinogen beta chain) and C4BPA (complement component 4 binding protein, alpha). Conclusion: This study suggests that OCD lesions may result from a cartilage defect whereas OC lesions may be triggered by both bone and cartilage defects, suggesting that different molecular mechanisms responsible for the equine osteochondrosis lesion subtypes and predisposition could be due to a defect in both bone and cartilage. This study will contribute to refining the definition of OC(D) lesions and may improve diagnosis and development of therapies for horses and other species, including humans

Endurance Exercise Ability in the Horse: A Trait with Complex Polygenic Determinism

Endurance horses are able to run at more than 20 km/h for 160 km (in bouts of 30–40 km). This level of performance is based on intense aerobic metabolism, effective body heat dissipation and the ability to endure painful exercise. The known heritabilities of endurance performance and exercise-related physiological traits in Arabian horses suggest that adaptation to extreme endurance exercise is influenced by genetic factors. The objective of the present genome-wide association study (GWAS) was to identify single nucleotide polymorphisms (SNPs) related to endurance racing performance in 597 Arabian horses. The performance traits studied were the total race distance, average race speed and finishing status (qualified, eliminated or retired). We used three mixed models that included a fixed allele or genotype effect and a random, polygenic effect. Quantile-quantile plots were acceptable, and the regression coefficients for actual vs. expected log10p-values ranged from 0.865 to 1.055. The GWAS revealed five significant quantitative trait loci (QTL) corresponding to 6 SNPs on chromosomes 6, 1, 7, 16, and 29 (two SNPs) with corrected p-values from 1.7 × 10−6 to 1.8 × 10−5. Annotation of these 5 QTL revealed two genes: sortilin-related VPS10-domain-containing receptor 3 (SORCS3) on chromosome 1 is involved in protein trafficking, and solute carrier family 39 member 12 (SLC39A12) on chromosome 29 is active in zinc transport and cell homeostasis. These two coding genes could be involved in neuronal tissues (CNS). The other QTL on chromosomes 6, 7, and 16 may be involved in the regulation of the gene expression through non-coding RNAs, CpG islands and transcription factor binding sites. On chromosome 6, a new candidate equine long non-coding RNA (KCNQ1OT1 ortholog: opposite antisense transcript 1 of potassium voltage-gated channel subfamily Q member 1 gene) was predicted in silico and validated by RT-qPCR in primary cultures of equine myoblasts and fibroblasts. This lncRNA could be one element of the cardiac rhythm regulation. Our GWAS revealed that equine performance during endurance races is a complex polygenic trait, and is partially governed by at least 5 QTL: two coding genes involved in neuronal tissues and three other loci with many regulatory functions such as slowing down heart rate

Chondrocytes Play a Major Role in the Stimulation of Bone Growth by Thyroid Hormone

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MOESM1 of A missense variant in the coil1A domain of the keratin 25 gene is associated with the dominant curly hair coat trait (Crd) in horse

Additional file 1: Table S1. List of animals included in the study. This table provides detailed information about identification, pedigree, phenotypes and genotypes of animals included in the study. The full pedigree is also provided. Table S2a. Details on markers and haplotypes in the mapped region. This table provides markers and their position, including the KRT25 variant along ECA11. Colors are used to depict haplotypes, assuming 18 founder haplotypes (12 “non-curly” haplotypes carried by crossbreed animals as well as the six most frequent other haplotypes, including one “curly” haplotype). Haplotypes are given for each animal, according to pedigree data. Table S2b. Delineation of the critical mapping region by haplotype analysis. Haplotypes were sorted out according to their sequence and the associated coat phenotype. Critical recombination events are easily detected by color changes and help identify the upper and lower bounds of the mapping interval. Table S3. PCR primers used to genotype candidate genes. This table provides detailed information about chromosome positions, alleles, gene names, ID, primer sequences, fragment lengths and Tm. Table S4. Statistics on genome-wide detection of functional variants. Table S4 provides statistics on variant detection based on NGS sequencing, with numbers of new and known variants (whole genome and genes) according to their consequence as predicted by VEP. Positional candidate variants are also reported. Table S5. List of gene variants predicted to impact protein functionality. Table S5 includes chromosome positions, alleles, gene ID and functional annotations. The last column (Existing variation) indicates variants; which were already known in dbSNP. Table S6. Concordant variants identified in the critical mapping interval. Table S6 includes chromosome positions, alleles, functional annotations and gene ID for concordant variants. Table S7. Horses genotyped by PCR and Sanger sequencing. Horse names, phenotype and genotype for the KRT24:g.21932167G>T, Top2A:g.22186465C>T and Top2A: g.22191762G>T variants are presented. Table S8. Pedigree information for the progeny of Walker’s Prince T and Dravkvallons Ite O Maguzu. Table S8 provides pedigree and phenotype data obtained from breeders and the Curly horse pedigree database ( http://www.curlyhorses.info/mainsearch.asp ). Table S9. Comparison of NCBI and Ensembl annotations within the keratin cluster on ECA11. Annotation features are listed along ECA11, from position 2,136,900 to 2,197,100 pb; some discrepancy is observed upstream of the critical mapping interval