• Asian-Australasian Journal of Animal Sciences
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• v.29 no.7
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• pp.925-937
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• 2016

In the last few decades, transgenic animal technology has witnessed an increasingly wide application in animal breeding. Reproductive traits are economically important to the pig industry. It has been shown that the bone morphogenetic protein receptor type IB (BMPR1B) A746G polymorphism is responsible for the fertility in sheep. However, this causal mutation exits exclusively in sheep and goat. In this study, we attempted to create transgenic pigs by introducing this mutation with the aim to improve reproductive traits in pigs. We successfully constructed a vector containing porcine BMPR1B coding sequence (CDS) with the mutant G allele of A746G mutation. In total, we obtained 24 cloned male piglets using handmade cloning (HMC) technique, and 12 individuals survived till maturation. A set of polymerase chain reactions indicated that 11 of 12 matured boars were transgene-positive individuals, and that the transgenic vector was most likely disrupted during cloning. Of 11 positive pigs, one (No. 11) lost a part of the terminator region but had the intact promoter and the CDS regions. cDNA sequencing showed that the introduced allele (746G) was expressed in multiple tissues of transgene-positive offspring of No.11. Western blot analysis revealed that BMPR1B protein expression in multiple tissues of transgene-positive $F_1$ piglets was 0.5 to 2-fold higher than that in the transgene-negative siblings. The No. 11 boar showed normal litter size performance as normal pigs from the same breed. Transgene-positive $F_1$ boars produced by No. 11 had higher semen volume, sperm concentration and total sperm per ejaculate than the negative siblings, although the differences did not reached statistical significance. Transgene-positive $F_1$ sows had similar litter size performance to the negative siblings, and more data are needed to adequately assess the litter size performance. In conclusion, we obtained 24 cloned transgenic pigs with the modified porcine BMPR1B CDS using HMC. cDNA sequencing and western blot indicated that the exogenous BMPR1B CDS was successfully expressed in host pigs. The transgenic pigs showed normal litter size performance. However, no significant differences in litter size were found between transgene-positive and negative sows. Our study provides new insight into producing cloned transgenic livestock related to reproductive traits.

• Journal of Veterinary Clinics
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• v.31 no.3
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• pp.258-261
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• 2014

A 4-week-old male piglet being maintained in a research facility was found dead without any previous clinical signs. The piglet had been born from a surrogate mother after somatic nuclear transfer as part of a xenotransplantation study. Ovaries for nuclear transfer were obtained from a private farm outside the research facility. Histopathologically, multifocal to coalescing granulomatous myocarditis was observed in the heart, characterized by infiltration of lymphocytes, macrophages and multinucleated giant cells, and by myocardial necrosis and fibrosis. Lymphoid tissues showed marked lymphoid depletion with infiltration by histiocytes or giant cells. Immunohistochemistry showed PCV-2 antigens in necrotic myocytes, macrophages and multinucleated giant cells in the heart, as well as in macrophages and giant cells in lymphoid depleted areas of lymphoid tissues. Reproductive failure associated with PCV-2 in aborted or stillborn piglets is frequently characterized by myocarditis, and similar lesions were observed in this 4-week-old piglet with PCV-2 infection. The PCV-2 infection in this piglet may have been due to contamination or infection of an ovary from the pig farm.

• Asian-Australasian Journal of Animal Sciences
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• v.24 no.5
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• pp.670-677
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• 2011

The growth rate of the young pig is generally much less than it's potential and may be constrained by endocrine status as well as nutrient intake. Growth factors are present in relatively high quantities in colostrum and play an important part in gut development. It is possible that supplementation of colostrum protein isolate may stimulate gut and whole body growth in the pig. Eight male and 8 female (Large Whitex${\times}$Landrace) piglets were weaned at 1 d of age after each pig had obtained colostrum from their dam, and were trained to consume one of two liquid diets. The two diets were based on either a colostrum protein isolate (n = 4 males and 4 females) or whey protein concentrate (n = 4 males and 4 females) and were formulated to contain equal levels of crude protein and amino acids. Pigs were fed their diets ad libitum for 28 days after which time 12 pigs were euthanised and various tissues and organs weighed. Pigs were bled for IGF-I analyses at 21 and 28 days of age. Daily gain was higher in pigs consuming the colostrum isolate (171 vs. 216 g/d, p = 0.010), particularly between 2 and 4 weeks of age (212 vs. 298 g/d, p = 0.010). Pigs tended to consume more of the liquid feed containing colostrum isolate (25.5 vs. 29.1 kg, p = 0.074) and gained more live weight per unit of liquid feed (0.203 vs. 0.223 g/g, p = 0.056). There were no effects of sex on growth performance. Pigs consuming the diet supplemented with colostrum isolate had higher (p<0.05) full gut weight (445 vs. 554 g, p = 0.026), empty gut weight (356 vs. 463 g, p = 0.008), stomach weight (42.2 vs. 54.4 g, p = 0.001), small intestine weight (222 vs. 275 g, p = 0.025) and large intestine weight (63.7 vs. 98.0 g, p = 0.005). Plasma IGF-I (99 vs. 150 ng/ml, p<0.001) and IGF-II (265 vs. 406 ng/ml, p<0.001) were higher in pigs fed colostrum isolate. Pigs consuming colostrum protein isolate ate more, grew faster and had higher plasma IGF-I concentrations than pigs consuming a diet with similar macronutrient content but devoid of growth factors.

• Journal of Embryo Transfer
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• v.32 no.3
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• pp.209-220
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• 2017

The estrogen-mediated effect of mesenchymal stem cells (MSCs) is a highly critical factor for the clinical application of MSCs. However, the present study is conducted on MSCs derived from adult donors, which have different physiological status with steroid hormonal changes. Therefore, we explores the important role of $17{\beta}$-estradiol (E2) in MSCs derived from female and male newborn piglets (NF- and NM-pBMSCs), which are non-sexually matured donors with steroid hormones. The results revealed that in vitro treatment of MSCs with E2 improved cell proliferation, but the rates varied according to the gender of the newborn donors. Following in vitro treatment of newborn MSCs with E2, mRNA levels of Oct3/4 and Sox2 increased in both genders of MSCs and they may be correlated with both estrogen receptor ${\alpha}$ ($ER{\alpha}$) and $ER{\beta}$ in NF-pBMSCs, but NM-pBMSCs were only correlated with $ER{\alpha}$. Moreover, E2-treated NF-pBMSCs decreased in ${\beta}$-galactosidase activity but no influence on NM-pBMSCs. In E2-mediated differentiation capacity, E2 induced an increase in the osteogenic and chondrogenic abilities of both pBMSCs, but adipogenic ability may increased only in NF-pBMSCs. These results demonstrate that E2 could affect both genders of newborn donor-derived MSCs, but the regulatory role of E2 varies depending on gender-dependent characteristics even though the original newborn donors had not been affected by functional steroid hormones.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.20 no.9
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• pp.306-314
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• 2019

As dipeptidyl peptidase-4(DPP-4) inhibitors are used widely as a secondary treatment for type 2 diabetes because they tend to be well tolerated with minimal side effects, the human DPP-4(hDPP-4) gene was injected into a pig zygote through micro-injection, and 1-cell stage fertilized embryos were then transplanted surgically into the oviduct. Three pigs were fertilized with hDPP-4 genes and produced sixteen piglets, in which one male piglet was identified to be transgenic. Finally, transgenic pigs showing hDPP-4 gene expression in the tail were produced. Western blot and RT-PCR analysis confirmed that the hDPP-4 is expressed strongly in the membrane cells of the transgenic pig, and that the hDPP-4 gene appears in various tissues and tails. This suggests that the expression vector is normally expressed in transgenic pigs. These results are anticipated to be a model animal to check the endocrine function for insulin resistance that occurs in a hDPP-4 transgenic pig and to increase its value for use as a material in newly developed medicines.