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Ito T.,ZEN NOH National Federation of Agricultural Co operative Associations | Sendai Y.,Central Research Institute for Feed and Livestock | Yamazaki S.,Tokyo Medical University | Yamazaki S.,Japan Science and Technology Agency | And 7 more authors.
PLoS ONE | Year: 2014

Although severe combined immune deficiency (SCID) is a very important research model for mice and SCID mice are widely used, there are only few reports describing the SCID pig models. Therefore, additional research in this area is needed. In this study, we describe the generation of Recombination activating gene-1 (Rag-1)-deficient neonatal piglets in Duroc breed using somatic cell nuclear transfer (SCNT) with gene targeting and analysis using fluorescence-activated cell sorting (FACS) and histology. We constructed porcine Rag-1 gene targeting vectors for the Exon 2 region and obtained heterozygous/homozygous Rag-1 knockout cell colonies using SCNT. We generated two Rag-1-deficient neonatal piglets and compared them with wild-type neonatal piglets. FACS analysis showed that Rag-1 disruption causes a lack of Immunoglobulin M-positive B cells and CD3-positive T cells in peripheral blood mononuclear cells. Consistent with FACS analysis, histological analysis revealed structural defects and an absence of mature lymphocytes in the spleen, mesenteric lymph node (MLNs), and thymus in Rag-1-deficient piglets. These results confirm that Rag-1 is necessary for the generation of lymphocytes in pigs, and Rag-1-deficient piglets exhibit a T and B cell deficient SCID (T-B-SCID) phenotype similar to that of rodents and humans. The T-B-SCID pigs with Rag-1 deficiency generated in this study could be a suitably versatile model for laboratory, translational, and biomedical research, including the development of a humanized model and assessment of pluripotent stem cells. © 2014 Ito et al. Source


Kubota Y.,Ibaraki University | Kubota Y.,Central Research Institute for Feed and Livestock | Goto T.,Ibaraki University | Hagiya Y.,Ibaraki University | And 4 more authors.
Stress | Year: 2016

Social stress may precipitate psychiatric disorders such as depression, which is related to the occurrence of the metabolic syndrome, including obesity and type 2 diabetes. We have evaluated the effects of social stress on central and peripheral metabolism using a model of depression in mice. In the present study, we focused on coenzyme A (CoA) molecular species [i.e. non-esterified CoA (CoASH), acetyl-CoA and malonyl-CoA] which play important roles in numerous metabolic pathways, and we analyzed changes in expression of these molecules in the hypothalamus and liver of adult male mice (C57BL/6J) subjected to 10 days of subchronic mild social defeat stress (sCSDS) with ICR mice as aggressors. Mice (n = 12) exposed to showed hyperphagia- and polydipsia-like symptoms and increased body weight gain compared with control mice which were not affected by exposure to ICR mice (n = 12). To elucidate the underlying metabolic features in the sCSDS model, acetyl-CoA, malonyl-CoA and CoASH tissue levels were analyzed using the acyl-CoA cycling method. The levels of hypothalamic malonyl-CoA, which decreases feeding behavior, were not influenced by sCSDS. However, sCSDS reduced levels of acetyl-CoA, malonyl-CoA and total CoA (sum of the three CoA molecular species) in the liver. Hence, hyperphagia-like symptoms in sCSDS mice evidently occurred independently of hypothalamic malonyl-CoA, but might consequently lead to down-regulation of hepatic CoA via altered expression of nudix hydrolase 7. Future studies should investigate the molecular mechanism(s) underlying the down-regulation of liver CoA pools in sCSDS mice. © 2016 Taylor & Francis. Source


Hirose K.,National Federation of Agricultural Co operative Associations | Ito T.,National Federation of Agricultural Co operative Associations | Fukawa K.,National Federation of Agricultural Co operative Associations | Arakawa A.,Japan National Institute of Agrobiological Science | And 3 more authors.
Animal Science Journal | Year: 2014

We evaluated multiple effects of genetic variations of five candidate loci (LEP, LEPR, MC4R, PIK3C3 and VRTN) on four production traits (average daily weight gain (ADG); backfat thickness (BFT); loin eye muscle area (EMA); and intramuscular fat content (IMF)) in a closed nucleus herd of pure Duroc pigs. Polymorphisms in LEPR, MC4R and PIK3C3 had significant single gene effects on ADG and BFT. The additive genetic variance in ADG and BFT (16.99% and 22.51%, respectively) was explained by genetic effects of these three loci. No correlations were observed between the LEP genotype and production traits in this study. Although we detected marginally epistatic interactions between LEPR and PIK3C3 on the eye muscle area, there were no significant epistatic effects on any traits among all loci pairs. These results suggest that LEPR, MC4R, PIK3C3 and VRTN may independently influence growth rate and fat deposition. Furthermore, the statistical models for predicting the breeding values of each trait had the lowest Akaike's information criterion values when considering the effect of the MC4R, LEPR, PIK3C3 and VRTN genotype simultaneously. These results suggest that LEPR, MC4R, PIK3C3 and VRTN are useful markers for accurately predicting breeding values in Duroc pigs. © 2013 Japanese Society of Animal Science. Source


Sugiyama T.,Niigata University | Kusuhara S.,Niigata University | Chung T.K.,DSM Nutritional Products Asia Pacific | Yonekura H.,Central Research Institute for Feed and Livestock | And 2 more authors.
Animal Science Journal | Year: 2013

The principal objective of this experiment was to evaluate the effect of 25-hydroxy-cholecalciferol (25-OH-D3) on the development of osteochondrosis in 6- to 110-kg castrated male pigs. The growth rate and serum calcium and inorganic phosphate levels neither increased nor decreased in response to supplementation of 25-OH-D3. However, supplemental 25-OH-D3 significantly increased serum levels of 25-OH-D3 and 1α,25-hydroxy-cholecalciferol without any influence on bone mineral density. The 25-OH-D3-treated group had significant (P<0.05) reduced incidence of osteochondrotic lesions compared to the control group as evidenced by macroscopically examining the articular cartilage of the distal humerus (32.4% vs. 59.3%) and distal femur (47.1% vs. 87.5%). Likewise, supplemental 25-OH-D3 significantly reduced osteochondrotic lesions over the control when histologically examining humerus (20.6% vs. 43.8%) and femur (52.9% vs. 87.5%). The results of this experiment suggested that 25-OH-D3 supplementation in pig diets had a tendency to promote normal endochondral ossification, inhibit osteochondrosis progression and possibly regenerate destroyed cartilage tissue. © 2012 Japanese Society of Animal Science. Source


Uemoto Y.,National Livestock Breeding Center | Sato S.,National Livestock Breeding Center | Ohnishi C.,National Livestock Breeding Center | Hirose K.,Central Research Institute for Feed and Livestock | And 4 more authors.
Animal Science Journal | Year: 2010

Leg weakness in pigs is a serious problem in the pig industry. We performed a whole genome quantitative trait locus (QTL) analysis to find QTLs affecting leg weakness traits in the Landrace population. Half-sib progeny (n = 522) with five sires were measured for leg weakness traits. Whole genome QTL mapping was performed using a half-sib regression-based method using 190 microsatellite markers. No experiment-wide significant QTLs affecting leg weakness traits were detected. However, at the 5% chromosome-wide level, QTLs affecting leg weakness traits were detected on chromosomes 1, 3, 10 and 11 with QTL effects ranging from 0.07 to 0.11 of the phenotypic variance. At the 1% chromosome-wide level, QTLs affecting rear feet score and total leg score were detected on chromosomes 2 and 3 with QTL effects of 0.11 and 0.13 of the phenotypic variance, respectively. On chromosome 3 and 10, some QTLs found in this study were located at nearby positions. The present study is one of the first reports of QTLs affecting fitness related traits such as leg weakness traits, that segregate within the Landrace population. The study also provides useful information for studying QTLs in purebred populations. © 2009 The Authors. Journal compilation © 2009 Japanese Society of Animal Science. Source

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