• Journal of Embryo Transfer
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• v.31 no.1
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• pp.65-72
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• 2016

This study was conducted to examine effects of mating type on the growth traits in an $F_2$ population produced by reciprocal intercrosses between Landrace and the Jeju Native Black pig (JBP). The $F_2$ progeny were produced by two different mating types based on the grand dams of founder breeds JBP (Cross_1) and Landrace (Cross_2). The body weights at 21 days after birth (BW21D) was significantly different between Cross_1 and Cross_2 (P<0.05), showing that the BW21D of Cross_1 has about 0.25 kg heavier than Cross_2. The significant differences were found between males and females for the growth traits including the body weights (BWB, BW21D, BW70D and BW140D) and average daily gains (ADG, eADG and lADG) (P<0.05). Males were heavier BWB, BW21D and BW140D levels, and higher ADG and lADG levels than females. On the other hand, females had heavier BW70D and higher eADG levels than those of males. When considering the mating types and sex simultaneously the Cross_2 males had the heaviest BW140D among the combinations of cross and sex. In conclusion, it is desirable to choose Landrace as grand dams in the reciprocal intercrosses between Landrace and JBP for producing their progeny construction and to plan the production of $F_2$ males for industrial purposes. These results suggested that it may be one of useful strategies to improve the productivity through out selection of the mating type of founder breeds and the progeny sex, especially in Landrace, JBP and their related populations.

• Journal of Embryo Transfer
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• v.33 no.4
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• pp.195-203
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• 2018

Interferon-tau (IFNT) is known as a major conceptus protein that signals the process of maternal recognition of pregnancy in ruminants. Also, multiple interferon genes exist in cattle, However, molecular mechanisms of these bovine IFNT (bIFNT) genes whose expressions are limited have not been characterized. We and others have observed that expression levels of bovine subtype IFNT genes in the tissues of ruminants; thus, bIFNT1 and other new type I (bIFNTc1/c2/c3) gene co-exist during the early stages of conceptus development and non-trophoblast cells. Its genes transcription could be regulated through CDX2 and ETS2 and JUN and/or cAMP-response element binding protein (CREB)-binding protein (CREBBP) expression, a transcription factor implicated in the control of cell differentiation in the trophectoderm. Bovine ear-derived fibroblast cells, were co-transfected with luciferase reporter constructs carrying upstream (positions -1000 to +51) regions of bIFNT1 and other new type I gene and various transcription factor expression plasmids. Compared to each - 1kb-bIFNT1/c1/c2/c3-Luc increased when this constructs were co-transfected with CDX2, ETS2, JUN and/or CREBBP. Also, Its genes was had very effect on activity by CDX2, either alone or with the other transcription factors, markedly increased luciferase activity. However, the degree of transcriptional activation of the bIFNTc1 gene was not similar to that bIFNT1/c2/c3 gene by expression plasmid. Furthermore, Sequence analyses also revealed that the expression levels of bIFNT1/c2/c3 gene mRNAs expression were highest on day 17, 20 and 22 trophoblast and, Madin-Darby bovine kidney (MDBK), Bovine ear-derived fibroblast (EF), and endometrium (Endo) non-trophoblast cells. But, bIFNTc1 mRNA had not same expression level, bIFNTc1 lowest levels than those of IFNT1/c2/c3 gene in both trophoblast and non-trophoblast cells. These results demonstrate that bovine subtype bIFNT genes display differential, in the trophoblast and non-trophoblast cells.

• Korean Journal of Animal Reproduction
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• v.20 no.4
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• pp.443-451
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• 1997

These studies were conducted to determine the sex of preimplantation Hanwoo embryos produced in vitro using polymerase chain reaction(PCR). Y chromosome specific and bovine speicific DNA primers were synthesized and tested for embryo sexing. Bovine IVF embryos were produced in TCM 199 and CR1aa medium, and classified by developmental stages on Day 7 to 9. The effects of developmental rates to bovine IVF blastocysts on sex ratio were also investigated using PCR methods. The results obtained in this study were as follows; 1. Developmental rates to blastocyst from IVM/IVF embryos in TCM 199 and CR1aa medium for 9 days were 23.5 and 30.2%, respectively, and there was significant difference between the media(P<0.05). 2. Male to female ratio of early, mid, expanded and hatching balstocyst produced on Day 7 were 0.7:1, 1.4:1, 2.2:1, and 2.5:1, respectively, and male embryos was significantly higher proportion in expanding and hatching blastocysts(P<0.01). 3. On Day 8, male to female ratio of early, mid, expanded and hatching blastocysts were 0.6:1, 1:1, 2.5:1, and 2.7:1, respectively. Both expanded and hatching blastocysts obtained a significantly higher proportion of males(P<0.01). 4. The male : female ratio of early, mid, expanded and hatching blastocyst produced on Day 9 was 0.6:1, 0.8:1, 1:1, and 2.2:1, respectively. Hatching blastocysts had a significantly higher ratio of males(P<0.01). The developmental rate of IVM/IVF embryos to blastocyst for 9 day culture was higher in CR1aa than that in TCM 199 medium. For the sex ratio by developmental stages of IVF embryos, male ratio was higher in expanded blastocyst but female in early blastocysts.

• Asian-Australasian Journal of Animal Sciences
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• v.22 no.4
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• pp.483-491
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• 2009

In vitro produced porcine embryos have potential application in reproductive biotechnology. However, their development potential has been very low. This study evaluated the in vitro developmental ability and quality of cloned and parthenogenetic porcine embryos having 2-4 cells or 5-8 cells on Day 2 of in vitro culture. Analysis of results showed that 2 to 4 cell embryos had higher ability to form blastocysts than 5 to 8 cell embryos (p<0.05). Blastocysts produced from culture of 2 to 4 cell embryos also contained higher cell numbers and had lower BAX:BCLxL transcript ratio than those produced from 5 to 8 cell embryos (p<0.05), thereby suggesting 2 to 4 cell embryos have higher development potential. Further investigation revealed that 5 to 8 cell embryos had higher incidence (100${\pm}$0.0%) of blastomeric fragmentation than 2 to 4 cell embryos (15.2${\pm}$5.5% for parthenogenetic and 27.7${\pm}$7.1% for cloned embryos). This suggests that low development potential of 5 to 8 cell embryos was associated with blastomeric fragmentation. In conclusion, we have shown that morphological selection of embryos based on cell number on Day 2 of in vitro culture could offer a practical and valuable non-invasive means to select good quality porcine embryos.

• Journal of Embryo Transfer
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• v.20 no.3
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• pp.303-309
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• 2005

This study was conducted to examine the effects of OA on metaphase of meiosis II and the mitochondrial activity of cytoplasm in bovine cumulus oocytes complexes(COCs) during in vitro maturation. Hanwoo COCs were collected from the slaughterhouse cow ovaries and matured in TCM199 supplemented with $0.1\%$ PVA, 0.2 uM, 2 uM, 20 uM OA for the maturation rate of OA concentration. For the maturation effects between OA and cycloheximide(CX), COCs were matured in TCM199 with 25 ug/mL CX, 25 ug/mL CX (6 hrs culture) plus 2 uM OA or 2 uM OA only at a atmosphere $5\%\;CO_2,\;95\%$ air $39^{\circ}C$ for 6, 12, 24 hrs. To evaluate the nuclear types of matured COCs, cumulus cells were removedby $0.5\%$ hyaluronidase sol. and oocytes were fixed in 1:3 acetic acid ethyl alcohol for 30 sec. and then stained with $0.1\%$ basic Fuchsin sol. For the detection of fluoriscent intensity (FI) of matures oocytes, cumulus cells were removed same as performed above and were stained with 20 nM mite tracker for 20 min. at $39^{\circ}C$. Mitochondrial activity of FI in matured oocytes was imaged by laser conforcal microscopy (Fluoview, Olympus, Japan) and were measured scanned face on 5 um from median to endpoint of oocytes. Statical analysis of nuclear types observed the three replicates was carried out with ANOVA and Fisher's protected least significant difference test using the STATVIEW program. FI of matures oocytes was compared the multiples of the least intensity among the measured oocytes. Maturing in TCM199 supplemented with $0.1\%$ PVA, 0.2 uM, 2 uM, 20 uM OA, metaphase B were showed 72.0, 50.0, 70.0, $68.8\%$, respectively and there were different significant(p<0.05). In the case of treatment with OA and CX, metaphase were $73.8\%,\;8.2\%,\;45.5\%,\;73.7\%$ in $0.1\%$ PVA-TCM199, 25 ug/mL CX, 25 ug/mL CX plus OA or 2uM OA only, respeclively. FI was revealed the increasing tendency during the process of maturation. Whereas FI in CX was decreased about 3 times compared to the other treatments of 6 hrs maturation. We conclude that OA regulates bovine COCs maturation and induces the mitochondrial activity during the process of maturation.

• Journal of Embryo Transfer
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• v.20 no.3
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• pp.239-253
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• 2005

Serum concentrations of Hanwoo steers and bulls as possible indicators of beef quality were analyzed to estimate their correlations with carcass traits. Blood samples were taken 2 months and right before shipping to abattoir and at the time of slaughter. And phenotypic correlation coefficients between serum concentrations and carcass traits were estimated. Beef yield index of steers was positively correlated with serum concentrations of total Protein (0.23), albumin (0.26), and calcium (0.31). But it was negatively co..elated with BUN (-0.30). Loin eye area was positively correlated with BUN (0.17) or with globulin (0.16). Back fat thickness was positively correlated with BUN (0.42) and inorganic phosphorus (0.20) being negatively correlated with total protein (-0.23), albumin (-0.33) and calcium (-0.33). Marbling score in the scale of 1 (scarcely marbled) through 9 (extremely marbled) was positively correlated with BUN (0.28) and negatively with IGF-I and calcium concentrations. Phenotype correlation coefficient of loin eye area with total protein concentration in the serum taken from steers right before shipment was estimated to be -0.16 and that with BUN was estimated to be -0.15. Serum concentrations of IGF, glucose, creatinine and on organic phosphorus from steers measured right before shipment were negatively correlated with respective correlation coefficient estimates as -0.21, -0.21, -0.19 and -0.18. Marbling score was negatively co..elated with serum creatinine (-0.16) measured at that time. Beef yield index of steers was positively correlated (0.31) with age adjusted calcium concentration in the serum taken at the time of slaughter. Correlation between body weight and BUN at slaughter was 0.17 At slaughter, loin eye area was negatively correlated with albumin (-0.19) and back fat thickness was also negatively correlated with age adjusted calcium concentration (-0.38). Marbling was negatively correlated with age adjusted calcium concentration(-0.17). Serum concentrations of testosterone, calcium and inorganic phosphorus taken in 2 months before slaughter were negatively but highly correlated with yield index(0.71, 0.67 and -0.71), respectively. Body weight at slaughter was positively was negatively correlated (0.67) with calcium level while dressing percentage was negatively (-0.69) correlated with serum glucose concentration, 2 months prior to slaughter. Correlation coefficients between back fat thickness and cortisol, between back fat thickness and inorganic phosphate were both positive (0.29 and 0.69). Marbling score was negatively correlated with creatinine (-0.81) and positively with BUN (0.87). Body weight loss during shipping was positively correlated with albumin and inorganic phosphate (0.77, 0.83). Yield index of bulls was positively correlated with serum testosterone concentration (0.66). Dressing percentage was positively and highly correlated with globulin (0.73). Back fat thickness of bulls, however, was negatively correlated with testosterone (-0.60). Loin eye area of bull carcasses was positively correlated with testosterone (0.40). Mar-blaine was negatively co..elated with creatinine (-0.55). Yield index of bulls and age adjusted HDLC concentration at slaughter was negatively correlated (-0.71). Dressing percentage of bulls was positively and highly correlated with globulin concentration (0.70). Back fat thickness was also positively correlated with HDLC (0.69) in the serum taken at slaughter. Correlation coefficients between carcass weight and triglyceride, between loin eye are and testosterone and between marbling score and creatinine or glucose were 0.51, -0.91 and -0.58, respectively.

• Journal of Embryo Transfer
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• v.22 no.3
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• pp.185-190
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• 2007

The objective of this study was to investigate the characteristics of various estrous behavior and ovulation time in dairy cows and heifers. In total, 73 ovulations and 61 estrous detection were observed in 89 Holstein cows. Various estrous behavior were observed during 72 hours from two days after $PGF_2{\alpha}$ injection and their relation with the time of ovulation(ultrasound examinations at 4-h intervals) was investigated. In estrous periods, the rate of sniffing, chin resting, mounting and standing heat was 81%, 78%, 78% and 56%, respectively in cows. In heifers, the rate of sniffing, chin resting, mounting and standing heat was 61%, 68%, 82% and 76%, respectively. Ovulation in cows and heifers occurred $25.58{\pm}7.94\;and\;25.55{\pm}5.72h$ after onset of estrus, and $13.42{\pm}7.14\;and\;7.48{\pm}7.41h$ after end of estrus, respectively. Interval between onset of estrus and ovulation time was significantly (p<0.05) shorter for standing heat ($17.33{\pm}5.83\;h$) than for mounting, sniffing and chin resting ($23.58{\pm}5.12\;h,\;24.25{\pm}6.09\;h,\;23.42{\pm}6.04\;h$) in cows but not significantly different in heifers. Interval between end of standing heat and ovulation time was significantly (p<0.05) shorter for heifer($6.38{\pm}4.80$) than for cows($13.05{\pm}4.53$). Our results show that characteristics of estrous behavior and ovulation in dairy heifers are different to that of cows.

• Journal of Embryo Transfer
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• v.20 no.3
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• pp.255-269
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• 2005

This study was aimed to investigate genetic relationships, variables, and correlations between economic traits and metabolic materials in serum components according to bleeding periods and breeding locations for the castrated and not castrated Hanwoo cattle at National Livestock Research Institute. Analysis of variance for serum hormones and metabolic materials showed significant differences by breeding locations except for testosterone and globulin. Statistical differences for serum components were detected by birth year except for cortisol, total protein, globulin and creatinine, and by castration except for total protein and BUN. All the serum components were tended to have sire effects except for testosterone resulting in some degree of additive gene actions. Breeding locations showed statistical significances for carcass weight and back fat thickness, but not in carcass rate, KPH, live weight and transportation weight loss. Effects of breeding locations and castration were significant for all weight measurement periods except for 9 month and 6 month, respectively. A significant sire effect was observed in all weight measurements. Least squared means for concentration of serum components by breeding year, season and castration were not significant. High concentration of cortisol, creatinine and triglyceride and low concentration of IGF-1 and glucose were detected in castrated cattle. Concentration of testosterone with castrated cattle was $5.2\%$ corresponding to non castrated cattle. Estimation of heritabilities of serum components using a sire model with restricted maximum likelihood were ranged 0.07 to 0.58. High heritabilities were estimated for total protein, albumin, globulin, cortisol, creatinine and BUN were 0.53, 0.54, 0.42, 0.45, 0.58 and 0.54, respectively. Low heritabilities were estimated fur calcium, testosterone and IGF-1 for 0.07, 0.15 and 0.12, respectively. Heritabilities for carcass weight, back fat thickness, meat yield index, KPH, and IMF were estimated as 0.39, 0.45, 0.30 0.13, and 0.93. Heritabilities of weights on 18, 12, 9, 6, and 24 month were estimated as 0.78, 0.76, 0.62, 0.58 and 0.58. Estimated heritabilities for average daily gain on 6${\~}$2, 12${\~}$18, and 18${\~}$24 month were 0.80, 0.75 and 0.19, respectively.

• Journal of Embryo Transfer
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• v.32 no.1
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• pp.1-8
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• 2017

Cryopreservation of germ cells from genetically proven animals could be a source of restoration tools from the risk of extinction or disappearance of wanted characteristics. Using frozen semen, the genetic gains of Korean native cattle have been increased greatly for 70 years. The preservation of genetic resources as a form of frozen semen straw has limited availability due to the numbers. To circumvent this weakness of frozen semen, we tested two re-freezing methods with different initial thawing temperatures using frozen Korean proven semen and rare breed semen from albino, black and chikso breeders. It has been known that human sperm could resist to cryo-damages by repeated freeze-thaw cycles, but not for Korean proven bulls number (KPN) or for rare breeds. Total 7 frozen semem from brindled(2), black(1), Korean Albino(2) and KPN(1) bulls were used for our research. After thawing straws under $5^{\circ}C/2min$ or $37^{\circ}C/40sec$ with low temperature water bath and thermo jug, spermatozoa were re-diluted with triladyl diluents after first thawing and re-frozen. Sperm motilities were compared between animals and treated groups after re-thawing. Mean values of motility and viability of refrozen/thawed sperm for expansion of the number of straws were significantly higher in $5^{\circ}C$ than in $37^{\circ}C$ (P < 0.05). However, the activity of viable sperm thawed at $5^{\circ}C$ was significantly decreased before refreezing. It is estimated that re-freezing of frozen semen from rare Korean native cattle is possible with resistant properties of survived spermatozoa.

• Korean Journal of Animal Reproduction
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• v.22 no.4
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• pp.425-435
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• 1998

To improve the efficiency of nuclear transplantation in bovine, in this study the development in vitro of nuclear transferred (NT) embryos was compared by different activation regimens of the enucleated oocytes. The effect of developmental stage and culture system of donor nuclei on fusion and development in vitro of NT embryos were also evaluated. Oocytes were collected from Hanwoo ovaries obtained from slaughterhouse and matured in Ham's F-10 supplemented with hormones. After 20~22 h maturation, the oocytes were vortexed to be free from cumulus cells and subsequently their nucleus and the first polar body were removed. Enucleated oocytes were divided into 3 groups for activation; the oocytes of group I were activated with ionomycin for 5 min and subsequently incubated in 6-dimetylarninopurine (DMAP) for 4 h, Those of group II were treated with DMAP for 4 h at 39 h after onset of in vitro maturation (IVM) and those of group III were kept in room temperature ($25^{\circ}C$) for 3 h at 39 h after onset of IVM. After in vitro fertilization (IVF) the embryos for muclear donor were cultured either by group culture (20 embryos /50 ${mu}ell$ drop) or individually (1 embryo /50 ${mu}ell$ drop) for 4 day and 5 day. At day 4 and 5 after IVF, blastomeres were separated in calcium-magnesium free medium, and then classified into small (day 5: $\leq$ 38 ${\mu}{\textrm}{m}$, day 4: $\leq$ 46 ${\mu}{\textrm}{m}$) and large (day 5 : $\geq$ 38 ${\mu}{\textrm}{m}$, day 4 ; $\geq$ 46 ${\mu}{\textrm}{m}$). The separated blastomeres were replaced into enucleated and activated recipient cytoplasm. The blastomere-oocyte complexes were fused by electrically. The NT embryos were cultured in TCM-199 containing 10% FCS in 39$^{\circ}C$, 5% $CO_2$ incubator for 7 day. The results obtained were summarized as follows; There were no differences in fusion and development to blastocyst between groups as group I (68%, 10%), group II (75%, 14%) and group III (73%, 9%), respectively. However, the cell number in blastocyst of NT embryos in group III were significantly fewer than in the other groups (P＜0.05). No differences in fusion and development to blastocyst were found between individual or group cultured and between small or large blastomeres of day 4 and day 5 donor embryos. From these results, it was concluded that the combination of ionomycin and DMAP, or treatment of DMAP at 39 h after onset of IVM were useful for the efficient of production of NT bovine embryos, and the individual cultured embryos could be simply used as donor nuclei for NT bovine embryo.