• Journal of Embryo Transfer
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• v.32 no.4
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• pp.287-295
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• 2017

Mitochondrial dysfunction is found in oocytes and transmitted to offspring due to maternal obesity. Treatment of obese mothers with endoplasmic reticulum (ER) stress inhibitors such as salubrinal (SAL) can reverse the mitochondrial dysfunction and result in normal embryonic development. Pig oocytes have also shown ER stress mostly in metaphase II stage. ER stress in oocytes may hinder the in vitro production of pig embryos. This study investigated the effect of ER stress inhibition by SAL treatment during in vitro maturation (IVM) of porcine oocytes at 1, 10, 50 and 100 nM concentrations. Firstly, we tested various concentrations of SAL. SAL at 10 nM showed higher (P < 0.05) developmental competence to the blastocyst stage (55.6%) after parthenogenesis (PA) than control (44.2%) while not different from other concentrations (49.2, 51.6, and 50.8% for 1, 50, and 100 nM, respectively). Secondly, we performed time-dependent treatment at 10 nM of SAL for IVM of oocytes. It revealed that treatment with SAL during 22 to 44 h of IVM significantly improved PA embryonic development to the blastocyst stage compared to control (40.5, 46.3, 51.7 and 60.2% for control, 0 to 22 h, 22 to 44 h and 0 to 44 h of IVM, respectively, P < 0.05). Glutathione (GSH) content is an indicator of cytoplasmic maturation of oocytes. Reactive oxygen species (ROS) have a harmful effect on developmental competence of oocytes. For this, we determined the intraoocyte levels of GSH and ROS after 44 h of IVM. It was found that SAL increased intraoocyte GSH level and also decreased ROS level (P < 0.05). Finally, we performed somatic cell nuclear transfer (SCNT) after treating oocytes with 10 nM SAL during IVM. SAL treatment significantly improved blastocyst formation of SCNT embryos compared to control (39.6% vs. 24.7%, P < 0.05). Our results indicate that treatment of pig oocytes with ER stress inhibitor SAL during IVM improves preimplantation development PA and cloned pig embryos by influencing cytoplasmic maturation in terms of increased GSH content and decreased ROS level in IVM pig oocytes.

• Asian-Australasian Journal of Animal Sciences
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• v.20 no.5
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• pp.768-774
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• 2007

To study the effect of dietary docosahexaenoic acid (DHA) enrichment on the expression of hepatic genes in pigs, weaned, crossbred pigs (30 d old) were fed diets supplemented with either 2% tallow or DHA oil for 18 d. Hepatic mRNA was extracted. Suppression subtractive hybridization was used to explore the hepatic genes that were specifically regulated by dietary DHA enrichment. After subtraction, we observed 288 cDNA fragments differentially expressed in livers from pigs fed either 2% DHA oil or 2% tallow for 18 d. After differential screening, 7 genes were found to be differentially expressed. Serum amyloid A protein 2 (SAA2) was further investigated because of its role in lipid metabolism. Northern analysis indicated that hepatic SAA2 was upregulated by dietary DHA enrichment (p<0.05). In a second experiment, feeding 10% DHA oil for 2d significantly increased the expression of SAA2 (compared to the 10% tallow group; p<0.05). The porcine SAA2 full length cDNA sequence was cloned and the sequence was compared to the human and mouse sequences. The homology of the SAA2 amino acid sequence between pig and human was 73% and between pig and mouse was 62%. There was a considerable difference in SAA2 sequences among these species. Of particular note was a deletion of 8 amino acids, in the pig compared to the human. This fragment is a specific characteristic for the SAA subtype that involved in acute inflammation reaction. Similar to human and mouse, porcine SAA2 was highly expressed in the liver of pigs. It was not detectable in the skeletal muscle, heart muscle, spleen, kidney, lung, and adipose tissue. These data suggest that SAA2 may be involved in mediation of the function of dietary DHA in the liver of the pig, however, the mechanism is not yet clear.

• Journal of Animal Reproduction and Biotechnology
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• v.37 no.1
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• pp.34-41
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• 2022

In vivo oocytes grow and mature in ovarian follicles whereas oocytes are matured in vitro in plastic culture dishes with a hard surface. In vivo oocytes show a superior developmental ability to in vitro counterparts, indicating suboptimal environments of in vitro culture. This study aimed to evaluate the influence of an agarose matrix as a culture substrate during in vitro maturation (IVM) on the development of pig oocytes derived from small antral follicles (SAFs). Cumulus-oocyte complexes (COCs) retrieved from SAFs were grown in a plastic culture dish without an agarose matrix and then cultured for maturation in a plastic dish coated without (control) or with a 1% or 2% (w/v) agarose hydrogel. Then, the effect of the soft agarose matrix on oocyte maturation and embryonic development was assessed by analyzing intra-oocyte contents of glutathione (GSH) and reactive oxygen species (ROS), expression of VEGFA, HIF1A, and PFKP genes, and blastocyst formation after parthenogenesis. IVM of pig COCs on a 1% (w/v) agarose matrix showed a significantly higher blastocyst formation, intra-oocyte GSH contents, and transcript abundance of VEGFA. Moreover, a significantly lower intra-oocyte ROS content was detected in oocytes matured on the 1% and 2% (w/v) agarose matrices than in control. Our results demonstrated that IVM of SAFs-derived pig oocytes on a soft agarose matrix enhanced developmental ability by improving the cytoplasmic maturation of oocytes through redox balancing and regulation of gene expression.

• Applied Microscopy
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• v.11 no.1
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• pp.39-50
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• 1981

An electron microscopical observation was carried out to compare the general shape of the mast cells and structures of granules inside the cells in the stomach of 5 species in 3 orders of Mammals. In convenience, the granules in the cytoplasm were abbreviated as follows: 1) Homogeneous granule, GR1 2) Particulate granule, GR2 a. Dark dense particulate granule, GR2-1 b. Less dense particulate granule, GR2-2 3) Reticular granule, GR5 a. Dark dense reticular granule, GR5-1 b. Light dense reticular granule, GR5-2 In Mammalia including goat, dog, cat, and hamster, most of cytoplasmic organelle were Golgi apparatus and mitochondria, and most of the cytoplasmic granules were highly densed GR1and GR2. However GR5-1 and GR5-2 appeared in guinea pig while one side sunken or crescent-like types occured in both dog and guinea pig. All mast cells were oval or spindle with cytoplasmic processes around the cell. There was also found vacuoles and vesicles in these cells. These results demonstrated that there was a morphological difference between species of vertebrate in the mast cells and their cytoplasmic granules. It was also suggested that a variety of structures of granules were closely related with the composition (histamine, heparin, serotonin, hyaluronic acid etc.) and mature of the granules.

• Proceedings of the Korean Biophysical Society Conference
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• 1996.07a
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• pp.39-39
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• 1996

Extracts were prepared from six different poisonous mushrooms in order to identify biologically active natural ligands. The effects of these extracts were examined on the microsomal $^{45}$ Ca$^{2+}$ uptake and $^{45}$ Ca$^{2+}$ release. Five out of six species apparently inhibited the activity of total microsomal ATPases prepared from the epithelial cells of pig airway. (omitted)ted)

• Food Science of Animal Resources
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• v.34 no.6
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• pp.799-807
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• 2014

Meat source fraud and adulteration scandals have led to consumer demands for accurate meat identification methods. Nucleotide amplification assays have been proposed as an alternative method to protein-based assays for meat identification. In this study, we designed Loop-mediated isothermal amplification (LAMP) assays targeting species-specific mitochondrial DNA to identify and discriminate eight meat species; cattle, pig, horse, goat, sheep, chicken, duck, and turkey. The LAMP primer sets were designed and the target genes were discriminated according to their unique annealing temperature generated by annealing curve analysis. Their unique annealing temperatures were found to be $85.56{\pm}0.07^{\circ}C$ for cattle, $84.96{\pm}0.08^{\circ}C$ for pig, and $85.99{\pm}0.05^{\circ}C$ for horse in the BSE-LAMP set (Bos taurus, Sus scrofa domesticus and Equus caballus); $84.91{\pm}0.11^{\circ}C$ for goat and $83.90{\pm}0.11^{\circ}C$ for sheep in the CO-LAMP set (Capra hircus and Ovis aries); and $86.31{\pm}0.23^{\circ}C$ for chicken, $88.66{\pm}0.12^{\circ}C$ for duck, and $84.49{\pm}0.08^{\circ}C$ for turkey in the GAM-LAMP set (Gallus gallus, Anas platyrhynchos and Meleagris gallopavo). No cross-reactivity was observed in each set. The limits of detection (LODs) of the LAMP assays in raw and cooked meat were determined from $10pg/{\mu}L$ to $100fg/{\mu}L$ levels, and LODs in raw and cooked meat admixtures were determined from 0.01% to 0.0001% levels. The assays were performed within 30 min and showed greater sensitivity than that of the PCR assays. These novel LAMP assays provide a simple, rapid, accurate, and sensitive technology for discrimination of eight meat species.

• Korean Journal of Veterinary Research
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• v.39 no.3
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• pp.513-522
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• 1999

DNA hybridization assay using probes prepared from liver was carried out to identify species characterization of the domestic animals. Gel electrophoresis showed that the target DNA extracted from raw muscle were 1kb and uniform pattern while fragments size of heated muscle were irregular. Hybridization was performed by adding 200ng/ml probe in hybridization solution and incubating for 12 hours at $68^{\circ}C$. To obtain good discrimination, applied washing buffer and washing step differently depending on the species. The probes of pig, horse and dog formed the specific hybrids with each target DNA respectively. Although cross reaction was detected in cattle, goat and sheep but signal intensity among these species made the discrimination possible each other. Such pattern was the same in the cases of chicken, turkey and duck. The hybridization pattern of heated muscle was similar to that of raw muscle in general, but the signal intensity was inferior to that of raw muscle. Species identification between closely related animal species, hybridized using the target DNA of such closely related animal species as a blocking agent, remarkable increase of discrimination from the evident decrease of non specific reaction compared with the control group. In addition, in the admixture where certain meat was included in the beef, pork, chicken meat, we could find whether any unjust meat was admixed or not. In this case, detection limit of certain meat in admixture was 1%.

• Korean Journal of Veterinary Research
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• v.29 no.2
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• pp.69-73
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• 1989

Nine strains of Yersinia species isolated from pigs and dogs in Korea, comprising 5 strains of Yersinia enterocolitica, 2 strains of Y kristensenii and each strain of Y pseudotuberculosis and Y intermedia, were examined for the presence of virulence-associated plasmids, calcium dependency and provocation of guinea pig conjuntivitis($Ser{\acute{e}}ny$ test). Three strains of Y enterocolitica isolated from pigs were positive in calcium dependency and harbored one plasmid of about 45 megadalton, but negative in $Ser{\acute{e}}ny$ test.

• Asian-Australasian Journal of Animal Sciences
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• v.16 no.7
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• pp.1046-1048
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• 2003

A polymerase chain reaction (PCR) assay was developed to differentiate buffalo meat from the meat of Ceylon spotted deer (Axis axis ceylonensis), Ceylon sambhur (Cervus unicolor unicolor), cattle (Bovine), goat (Caprine), pig (Porcine), and sheep (Ovine). A set of primers were designed according to the sequence of the mitochondrial cytochrome b gene of bubalus bubalis and by PCR amplification a band of approximately 242 bp band was observed with buffalo DNA. These primers did not cross-react with DNA of other animal species tested in the study under the specified reaction conditions. A band of 649 bp was observed for all animal species tested when DNA was amplified with the universal primers indicating the presence of mitochondrial DNA in the samples. The technique was sensitive enough to identify rotten (10 days post slaughter), dried and cooked buffalo meat. The absence of a cross reaction with human DNA using the buffalo specific primers eliminates possible false positive reactions.

• Reproductive and Developmental Biology
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• v.36 no.1
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• pp.79-85
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• 2012

Minipigs are regarded as one of the most important laboratory animal in that anatomical and physiological properties are similar to human and their reproduction efficiency is relatively higher compared to other large animal species. Particularly, several diseases that cannot be mimicked in rodent models are successfully occurred or induced in pig models therefore it has been interested in a valuable model for human diseases. Pigs are also 'standard' species in xenotransplantation research. To maximize experimental outcome using minipigs, establishment and management of proper animal facility, right animal husbandry and control of pathogens are very important. In this review, we summarized several international guidelines related with minipigs published by several companies or governments and discuss optimal conditions for providing informative ideas to the researchers who want to use minipigs in their future studies.