Genetic analysis of the hair-length of Sapsaree dogs, a Korean native dog breed, showed a dominant mode of inheritance for long hair. Genome-Wide Association Study (GWAS) analysis and subsequent Mendelian segregation analysis revealed an association between OXR1, RSPO2, and PKHD1L1 on chromosome 13 (CFA13). We identified the previously reported 167 bp insertion in RSPO2 3’ untranslated region as a causative mutation for hair length variations. The analysis of 118 dog breeds and wolves revealed the selection signature on CFA13 in long-haired breeds. Haplotype analysis showed the association of only a few specific haplotypes to the breeds carrying the 167 bp insertion. The genetic diversity in the neighboring region linked to the insertion was higher in Sapsarees than in other Asian and European dog breeds carrying the same variation, suggesting an older history of its insertion in the Sapsaree genome than in that of the other breeds analyzed in this study. Our results show that the RSPO2 3’ UTR insertion is responsible for not only the furnishing phenotype but also determining the hair length of the entire body depending on the genetic background, suggesting an epistatic interaction between FGF5 and RSPO2 influencing the hair-length phenotype in dogs.
We developed a high-resolution and comprehensive typing method for swine leukocyte antigen 3 (SLA-3), an MHC class I gene, employing locus-specific genomic PCR followed by subsequent direct sequencing. A total of 292 individuals from nine pure, one cross-breed and six cell lines were successfully typed. A total of 21 SLA-3 alleles were identified, of which four were found to be novel alleles. However, the allelic diversity of SLA-3 was lower than that of previously reported class I genes, SLA-1 and -2. More SLA-3 alleles were observed in the Landrace and Yorkshire breeds than the other breeds. SLA-3*04:01 was identified in seven out of nine breeds and was the most widely distributed allele across all breeds. Therefore, the typing method reported in this study completes our efforts to develop high-resolution typing methods for major SLA molecules, facilitating the combined analysis of major SLA genes from field samples, which is important to understand the relationship between the adaptive immune responses against pathogens and the immunogenetic makeup of an individual.
In an attempt to investigate the effect of Hymenolepis nana infection on immunological responses to sRBC in ICR strain of mice, cellular and humoral immune responses were chronologically monitored after sensitization with sRBC. Mice weighing about 20 g were allocated into artificial and natural infection groups. The shell-free eggs of H. nana were inoculated into mice on the day 0 (initial) and day 10 in the former group, and praziquantel (25 mg/kg/day) was administered for 3 days to the one half of the mice at the 15th day after the first inoculation and to all of the mice in natural infection group. In artificial infection group, the delayed-type hypersensitivity (DTH) to sRBC was considerably decreased on the day 10 after the first inoculation, and then elevated gradually to normal. Eosinophils in the peripheral blood increased slightly. The hemagglutinin (HA) and hemolysin (HE) titers during the early stage were shown to be more or less higher than those of control. Thereafter, the titers were returned to normal, followed by a transient decrease on the day 15 post-infection. The sRBC rosette and antibody-treated rosette-forming capacities on the day 15 post-infection were temporarily lowered but became higher thereafter. The mucosal mast cells (MMC) in the small intestine were gradually increased to make a peak on the day 10 post-infection and then maintained more or less at lower level. After praziquantel treatment, the DTH and the number of eosinophils were decreased slightly and the MMC number and sRBC rosette-forming capacity were considerably decreased. The titers of HA and HE and antibody-treated rosette-forming capacity, however, were elevated in general. In natural infection group, the DTH, the number of eosinophils, and MMC which were elevated due to H. nana infection were gradually returned to normal after praziquantel treatment. The titers of HA and HE which were decreased by parasite infection were increased to normal after the treatment. However, the capacities of sRBC rosette or antibody-treated rosette formation were maintained at low levels in spite of the treatment. These results revealed that the immune responses to sRBC were significantly activated during H. nana infection, although they were transiently decreased during the days 10-15 post-infection.
Immortalized cell lines are valuable resources to expand the molecular characterization of major histocompatibility complex genes and their presented antigens. We generated a panel of immortalized cell lines by transfecting human telomerase reverse transcriptase (hTERT) into primary fibroblast cells prepared from ear, fetal, and lung tissues of 10 pigs from five breeds and successfully cultured them for 30–45 passages. The cell growth characteristic of the immortalized fibroblasts was similar to that of primary fibroblast, which was unable to form colonies on soft agar. The genotypes of major swine leukocyte antigen (SLA) genes, including three classical class I (SLA-1, -2, and -3) and three class II genes (DQB1, DRB1, and DQA), were determined using high-resolution typing. A total of 58 alleles, including a novel allele for SLA-2, were identified. Each cell line was unique. A cell line derived from a National Institutes of Health miniature pig was homozygous across the six major SLA genes. The expression levels of SLA classical class I genes varied among the cell lines and were slightly upregulated in the immortalized compared to the primary cells based on semiquantitative reverse transcription polymerase chain reaction. The immortalized porcine fibroblast cell lines with diverse SLA haplotypes that were developed in this study have potential to be applied in studies regarding the molecular characteristics and genetic structure of SLA genes and epitope–major histocompatibility complex interactions in pigs.
Understanding the influence of genetic variations in olfactory receptor (OR) genes on the olfaction-influenced phenotypes such as behaviors, reproduction, and feeding is important in animal biology. However, our understanding of the complexity of the OR subgenome is limited. In this study, we analyzed 1,120 typing results of 20 representative OR genes belonging to 13 OR families on 14 pig chromosomes from 56 individuals belonging to seven different breeds using a sequence-based OR typing method. We showed that the presence of copy number variations, conservation of locus-specific diversity, abundance of breed-specific alleles, presence of a loss-of-function allele, and low-level purifying selection in pig OR genes could be common characteristics of OR genes in mammals. The observed nucleotide sequence diversity of pig ORs was higher than that of dogs. To the best of our knowledge, this is the first report on the individual- or population-level characterization of a large number of OR family genes in livestock species.
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