• Asian-Australasian Journal of Animal Sciences
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• 제20권5호
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• pp.615-621
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• 2007

AMP-activated protein kinase alpha 2 (PRKAA2) plays a key role in regulation of fatty acid and cholesterol metabolism. This study investigated the porcine PRKAA2 gene as a positional candidate for intramuscular fat and backfat thickness traits in pig chromosome 6. A partial fragment of the porcine PRKAA2 gene, amplified by PCR, contained a putative intron 3 including a part of exon 3 and 4, comparable with that of human PRKAA2 gene. Within the fragment, several single nucleotide polymorphisms were identified using multiple sequence alignments. Of these, TaqI restriction enzyme polymorphism was used for genotyping various pig breeds including Korean reference family. Using linkage and physical mapping, the porcine PRKAA2 gene was mapped in the region between microsatellite markers SW1881 and SW1680 on chromosome 6. Allele frequencies were quite different among pig breeds. The full length cDNA of the porcine PRKAA2 (2,145 bp) obtained by RACE containing 1,656 bp open reading frame of deduced 552 amino acids, had sequence identities with PRKAA2 of human (98.2%), rat (97.8%), and mouse (97.5%). These results suggested that the porcine PRKAA2 is a positional candidate gene for fat deposition trait at near telomeric region of the long arm of SSC 6.

• 약학회지
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• 제50권6호
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• pp.409-414
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• 2006

Although mammalian sperms are cryopreserved for in vitro fertilization a process of cryopreservation decreases the fertility. Acrosome reaction requires depolarization-induced Ca$^{2+}$ influx and Ca$^{2+}$ releases from the Ca$^{2+}$ stores. To examine whether the cellular Ca$^{2+}$ mobilization is altered by a sperm cryopreservation we compared cytosolic Ca$^{2+}$ signals between fresh and cryopreserved pig sperms using confocal Ca$^{2+}$ imaging. The magnitudes of depolarization induced Ca$^{2+}$ increases were significantly smaller in cryopreserved sperms. Exposures to 10 mM caffeine or 5 ${\mu}$M thapsigargin elicited less Ca$^{2+}$ increases in the cryopreserved sperms compared to fresh sperms. In addition, progesterone-trig-gered Ca$^{2+}$ rises, that are thought to enhance acrosome reaction, were completely abolished in the cryopreserved sperms. These results suggest that storage and(/or) release of Ca$^{2+}$ from the intracellular Ca$^{2+}$ stores in pig sperms are significantly impaired by the process of cryopreservation.

• Asian-Australasian Journal of Animal Sciences
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• 제17권5호
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• pp.612-616
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• 2004

This study was carried out to compare the semen characteristics, frozen-thawed sperm viability and testosterone concentration and in vitro fertilization (IVF) and development of in vitro matured pig oocytes between two Yorkshire boars. Semen and blood samples were collected once per week from October to November 2002 from two adult Yorkshire boars at 18 months of age with 170 kg body weight. Sperm were deep frozen in 5 ml maxi-straws with lactose-egg yolk and N-acetyl-D-glucosamine (LEN) diluent and stored in liquid nitrogen. Blood samples were obtained at 10 a.m. by inserting a 21 gauge, hypodermic needle attached to 10 ml syringe into surface veins in the ear. The concentration of testosterone was determined by Competitive Enzyme Immunoassay. Ovaries were collected from prepubertal gilts at a local slaughter house. Cumulus oocyte complexes were aspirated from antral follicles (3 to 6 mm in diameter). The medium used for oocyte maturation was modified TCM 199. After about 22 h of culture, oocytes were cultured without cysteamine and hormones for 22 h at $38.5^{\circ}C$, 5% $CO_2$ in air. For IVF, one frozen 5 ml straw was thawed at $52^{\circ}C$in 40 sec and was diluted with 20 ml Beltsville thawing solution at room temperature. Sperm were washed 2 times in mTLP-PVA and inseminated without preincubation after thawing. Oocytes were inseminated with $2{\times}10^7$/ml sperm concentration. Oocytes were coincubated for 6 h in 500 ${\mu}$l mTBM fertilization medium. At 6 h after IVF, oocytes were transferred into 500 ${\mu}$l NCSU-23 culture medium for further culture of 48 and 144 h. There were no significant differences in the semen volume, motility, normal acrosome morphology and sperm concentration of raw semen between A and B of Yorkshire boar. However, motility and normal acrosome of boar A were higher than those of boar B at 0.5, 2, 3, 4, 5 and 6 h incubations of frozen-thawed sperm. Testosterone concentration (3.75 ng/ml) of boar A was higher than that (2.34 ng/ml) of boar B. The rate of blastocyst formation (15.1%) of boar A was higher than that (10.4%) of boar B. In conclusion, serum testosterone concentration of boar showed very important role for the frozen-thawed sperm viability and the blastocyst formation of pig oocytes matured in vitro.

• Reproductive and Developmental Biology
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• 제35권1호
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• pp.23-31
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• 2011

Two piglets and one juvenile pig were used to investigate closely what types of cells express green fluorescent protein (GFP) and if any, whether the GFP-tagged cells could be used for stem cell transplantation research as a middle-sized animal model in bone marrow cells of recloned GFP pigs. Bone marrow cells were recovered from the tibia, and further analyzed with various cell lineage markers to determine which cell lineage is concurrently expressing visible GFP in each individual animal. In the three animals, visible GFP were observed only in proportions of the plated cells immediately after collection, showing 41, 2 and 91% of bone marrow cells in clones #1, 2 and 3, respectively. The intensity of the visible GFP expression was variable even in an individual clone depending on cell sizes and types. The overall intensities of GFP expression were also different among the individual clones from very weak, weak to strong. Upon culture for 14 days in vitro (14DIV), some cell types showed intensive GFP expression throughout the cells; in particular, in cytoskeletons and the nucleus, on the other hand. Others are shown to be diffused GFP expression patterns only in the cytoplasm. Finally, characterization of stem cell lineage markers was carried out only in the clone #3 who showed intensive GFP expression. SSEA-1, SSEA-3, CD34, nestin and GFAP were expressed in proportions of the GFP expressing cells, but not all of them, suggesting that GFP expression occur in various cell lineages. These results indicate that targeted insertion of GFP gene should be pursued as in mouse approach to be useful for stem cell research. Furthermore, cell- or tissue-specific promoter should also be used if GFP pig is going to be meaningful for a model for stem cell transplantation.

• 한국동물위생학회지
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• 제32권1호
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• pp.27-32
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• 2009

We studied the seroprevalence of four respiratory pathogens in Korean swine farms located in Chungnam, Chungbuk, Gyeongnam and Gyeongbuk provinces during the period of spring of 2007 to winter of 2008. Serological tests were performed using commercial ELISA kits. A total of 530 serum samples were tested for the antibodies against porcine reproductive and respiratory syndrome virus (PRRSV), porcine circovirus type 2 (PCV2), Mycoplasma hyopneumoniae (M. hyo) and Actinobacillus pleuropneumoniae (APP). Seroprevalence for four respiratory pathogens were estimated by ELISA-positive rates of the submitted samples. The overall seropositive rates of PRRSV, APP, M. hyo and PCV2 were 32.6%, 10.6%, 38.4% and 88.5%, respectively. By production stage, the seropositive rate for PRRSV was highest in nursery pig populations (46.2%). In contrast, the highest seropositive rates of APP and M. hyo were observed in sow and growing pigs. However, the seroprevalence of PCV2 was ranged from 85.7% to 89.6%, showing no significant difference among the production stages. In the seroprevalence by season, PRRSV, APP and M. hyo infections revealed typical seasonal patterns that the peaks of the seropositive rates were observed between early winter and late spring. In case of PCV2, no particular seasonal patterns were noticed. The pig herds in Gyeongbuk province where PMWS was endemic during the period of survey showed the highest seropositive rates for PRRSV (44.6%), M. hyo (47.5%), and PCV2 (92.7%). Seropositive rates for APP of four provinces were approximately 10%. These results might be valuable for control and prevention of the respiratory diseases and helpful to define strategies related to vaccine applications.

• BMB Reports
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• 제41권6호
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• pp.466-471
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• 2008

Three isoforms of pig PDE4B were cloned and classified as two forms: PDE4B1 and PDE4B3, which contain UCR1 and UCR2; and PDE4B2, which contains only UCR2. The amino acid sequences of each isoform showed good conservation in human and rat. PDE4B2 is expressed in a wide range of tissues, but PDE4B1 and PDE4B3 are not. Using an informative SNP for the Iberian x Landrace intercross detected from intron 12, a linkage map was constructed. The location of PDE4B was estimated at 123.6 cM outside of the QTL-CI (124-128 cM) for IMF. However, the QTL-CI for IMF was reconfirmed with high significance, and its position was narrowed down to an interval of 4 cM (the region defined by markers PDE4B and SW1881). Using radiation hybrid mapping, LEPR, LEPROT, DNAJC6, AK3L1 and AK3L2 were selected as positional and/or functional candidates related to the QTL.

• 생명과학회지
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• 제28권4호
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• pp.389-397
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• 2018

당 단백질에 붙어 있는 당사슬 구조는 형질전환 돼지 유즙으로 분비되는 의약용 단백질의 생물학적 활성, 안정성 그리고 안전성에 영향을 줄 수 있다. 형질전환 동물을 이용한 치료용 당 단백질 생산은 유선 세포에서 이루어지는 당사슬 부가능력에 의해 제한되며, 균일한 당사슬 형태를 가지는 당 단백질 생산은 도전 과제로 남아있다. ${\beta}$-1,3-N-acetylglucosaminylatransferase1 (B3GNT1) 유전자는 N-아세틸글루코사민에 갈락토오스 잔기를 부착시키는 단백질 당화기작에 중요한 효소이지만, 돼지 당 전이효소에 대한 정보는 매우 제한적이다. 따라서, 돼지 B3GNT1 (pB3GNT1) 유전자를 클로닝하고 N-아세틸글루코사민에 갈락토오스 잔기를 부착시키는 기능적 특성을 조사하였다. 몇가지 다른 프라이머를 사용하여 전체 전사영역(ORF)을 함유하는 부분적인 pB3GNT1 mRNA 염기서열을 간 조직으로부터 분리하였다. 클로닝 된 pB3GNT1의 ORF는 1,248개의 뉴클레오티드를 가지며, 415개 아미노산 잔기로 구성되어 있었다. pB3GNT1 유전자의 장기별 발현특성은 성돈 및 자돈의 여러 기관에서 분석하였다. pB3GNT1 mRNA 발현 수준은 심장, 소장 보다는 근육에서 높았지만 폐에서는 낮았다. pB3GNT1의 기능적 특성 분석을 위해 돼지 신장 세포주(PK-15)에서 pB3GNT1 유전자의 안정적인 발현을 확립하였다. 그 결과, PK-15 세포에서 pB3GNT1 발현에 의한 당화 패턴은 총 시알산 증가에는 영향을 미치지 않지만, poly-N-아세틸글루코사민은 증가하는 것으로 나타났다. 본 연구는 생물반응기로 형질전환 돼지를 이용할 때 희망하는 당사슬을 부가하여 치료 가능성을 높이며 개선된 활성을 나타내는 당단백질 생산에 도움이 될 것이다.

• 대한수의학회지
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• 제50권3호
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• pp.247-251
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• 2010

This study compared the instrument performance and tissue healing of a steel scalpel with a $CO_2$ laser in an animal urinary bladder surgery model. Landrace and Yorkshire mixed breed pigs were used. Two symmetrical incisions were made in urinary bladder of each pig. One incision was made on the left side of ventral aspect on urinary bladder using a steel scalpel, while the other incision was performed on the right side using a $CO_2$ laser with an 8W output power. Each instrument was evaluated clinically for speed, ease of incision, and extent of bleeding. At 7 and 21 days after initial wounding, each wound was taken for histological observations. The scalpel was an easier instrument to use in the confines of the urinary bladder tissue, compared with the laser. However, there was no significant difference between the two groups. The amount of bleeding was less in the laser group but the time of the incisions was shorter with the scalpel. Scalpel incisions showed complete restoration of the epithelium and muscularis. On the other hand, the laser incisions showed incomplete restoration of the epithelium and muscularis. However, most of wound healing in the laser incisions was accomplished according to the time lapse. Although the scalpel produced less damage to the urinary bladder tissue and was easier to handle than the $CO_2$ laser, it did not provide hemostasis that was helpful for use on highly vascular tissue. The $CO_2$ laser provided good hemostasis, but delayed wound healing. In conclusion, the $CO_2$ laser provided better hemostasis and better surgical field than the scalpel. The $CO_2$ laser was used effectively in urinary bladder incision.

• 한국임상수의학회지
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• 제24권2호
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• pp.114-118
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• 2007

The objective of this study was to investigate the effects of implanted chitosan applied to surgically created wound in pigs. Six healthy $2{\sim}3$ months old Landrace and Yorkshire mixed breeds of both genders were used. A 2 cm straight skin incision was made and undermined skin ($2{\times}2cm$) over on the each pig's both sides of dorsal midline at 0, 7. 14 and 18 days. One wound (left side) was implanted 0.4 mg of cotton type chitosan and other wound was treated saline (3 ml). Each wound was closed with two interrupted suture of 2-0 sutures. The wounds created at 18, 14.7 and 0 days were named post-wounding day (PWD) 3, 7, 14 and 21, respectively. At 21 days after initial wounding, each wound was taken for histological observations. Reepithelialization tended to be greater in the chitosan group than in the control group at PWD 3 and 14. Granulation tissue formation did not show especial differences in two groups. Number of inflammatory cells was lesser statistically in level in the chitosan group than those in the control group at 21 days after wounding (p<0.05). Fibroblasts and neovasculature tended to be greater in the chitosan group than in the control group at PWD 3 and 7, and tended to be lesser in the chitosan group than in the control group at PWD 14 and 21. Collagen and fibrin were observed to be evenly distributed around the wound in the chitosan group. But collagen and fibrin were observed to be converged along the wound in the control group.

• Reproductive and Developmental Biology
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• 제29권1호
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• pp.43-48
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• 2005

돼지 웅성 페르몬인 5a-androst-16-en-3-one을 대체할 수 있는 활성 분자를 탐색하여 가축의 생산과 수요를 조절하기 위한 생물학적 자극 통제 수단으로 활용하고자 냄새 분자로서 2-cyclohexyloxytetrahydrofurane (A) 및 2-phenoxytetrahydrofurane (B) 유도체들의 구조 변화와 수용체인 porcine odorant binding protein (pOBP)에 대한 결합친화력 상수(p[Od.]/sub 50/) 사이의 정량적인 구조-활성관계에 관한 분자 HQSAR 모델XI을 유도하였다. 냄새 분자 중에서 cyclohexyl-치환체(A)가 phenyl-치환체(B)보다 높은 결합 친화력을 나타내었으며(A>B) 모델XI은 분자 조각크기(5∼8), 홀로그램 길이(97 bin)의 키랄성(chirality)조건에서 예측성(q²=0.916)과 상관성(r²=0.988)이 매우 양호하였다. 기여도(contribution map)로부터 냄새 분자의 결합 친화력 상수에 기여하는 부분은 2-oxyfuryl group의 C3 및 C5 원자인 반면에 cyclohexyl 고리상 tert-butyl group의 중심 탄소 원자와 furyl group의 C4 원자 부분은 기여하지 않았다.