The objective of this study was to investigate the changes of oxidative stress and antioxidant enzyme during in vitro development with washing culture oil in porcine embryos. During the culture, the four types of culture oil such as paraffin oil with or without washing and mineral oil with or without washing were examined. The oil was washed with PZM-3 during 7 days and collected oil only. The embryos were stained with CellTrackerTM Red, DC-FDA and Hoechst 33342 to confirm the effects of the oil. As a results, Cleavage rates and total cell number were no difference among the four oil groups. However, ≥16 cell embryos were significantly different in fore type oil treat-ment and blastocyst rate was significantly higher washing paraffin treatment than in other group(p<0.05). Also, the expression of free radical were lower in washing paraffin oil than in other groups (p<0.05). On the other hand, the expression of glutathione were not significant different among paraffin oil with or without washing and mineral oil with or without washing, however washing paraffin oil and washing mineral groups were higher than other treat-ment groups. In conclusion, the washing oil was expected with positive effects on in vitro development in porcine embryos.
In this study, to further understand the mechanism of animal growth and to develop a miniature transgenic animal model, we constructed and tested tetracycline-inducible RNAi system using shRNA targeting the mRNA of mouse insulin-like growth factor (mIGF-1) or mouse growth hormone receptor (mGHR) gene. Quantitative real-time PCR analysis of mouse liver cell (Hepa1c1c7) cells transfected with these vectors showed 85% or 90% of expression inhi-bition effect of IGF-1 or GHR, respectively. In ELISA analysis, the protein level of IGF-1 in the cells expressing the shRNA targeting IGF-1 mRNA was reduced to 26% of non-transformed control cells. Unexpectedly, in case of using shRNA targeting GHR, the IGF-1 protein level was decreased to 75% of control cells. Further experiments are needed to explain the lower interference effect of GHR shRNA in IGF-1 protein. Accumulated knowledge of this approach could be applicable to a variety of related biological area including gene function study, gene therapy, development of miniature animals, etc.
Oral exposure of humans by excess amounts of arsenic may cause disturbances of the reproductive system. In the present study, such exposure was modelled in rats, with the support of sperm principal parameters and histopa-thological observations. Male Sprague–Dawley rats were randomly divided into three groups where the group I was served as a normal control, group II was received sodium meta-arsenite as arsenic (10 mg/kg b.w/day) and a combi-nation of sodium meta- arsenite and sodium selenite (3 mg/kg b.w/day) in group III. After 6 weeks, there was no significant change in testis weight and in total motility of all the three experimental groups, whereas, rapid moving spermatozoa, moderately moving spermatozoa and slow moving spermatozoa were significantly decreased in arsenic treated rats as compared to control rats. The other sperm principal parameters like progressiveness, average path velocity, straightness linear velocity (VSL), curvilinear velocity (VCL), straightness, linearity sperm head elongation ratio, area, linearity amplitude of lateral head department (ALH) and beat cross frequency (BCF) were found to be reduced in arsenic intoxicated rats. These results are not correlated with the histological studies. On oral admini-stration of selenium ameliorated the adverse effects of arsenic as compared to arsenic alone treated rats. Our findings clearly demonstrate that administration of selenium could prevent some of the deleterious effects of arsenic in the testis.
Humulus japonicus is an ornamental plant in the Cannabaceae family. Although the mode of action of Humulus japonicus is not fully understood, a strong relationship was observed between anti-inflammatory and anticancer in some types of cells. Recent studies also have shown that Humulus japonicus possesses anti-inflammatory activities and may significantly improve antioxidant potential in Raw 264.7 macrophage cells. Thus, the aim of this study was eva-luated the effect of Humulus japonicus extract on sperm motility and subsequent preimplantation developmental com-petence of the bovine embryos. After in vitro maturation, the oocytes with sperms were exposed in in vitro fertilization (IVF) medium supplemented with Humulus japonicus extract (0.01, 0.05, 0.1 μg/mL, respectively) for 1 day. In our results, exposure of IVF medium to Humulus japonicus extract did not affect sperm motility and percentage of pene-trated oocytes but ROS intensity was significantly decreased by 0.01 μg/mL compared with other groups (p< 0.05). Moreover, treatment with 0.01 μg/mL of Humulus japonicus extract was higher the frequency of blastocyst formation than the any other groups (p<0.05). Otherwise, treatment with 0.01 μg/mL of Humulus japonicus extract not increased the total cell number but reduced apoptotic-positive nuclei number. In conclusion, our results indicate that supple-mentation of Humulus japonicus extract in IVF medium may have important implications for improving early embryo-nic development in bovine embryos
This study was conducted for SNPs in the 5'-regions of estrogen receptor-α(ESR-α), and association with calving interval (CI), service per conception (SPC) and 305 days milk yield in Hanwoo and Holstein dairy cattle. The genet-ic improvement was incurred low reproduction performance. The objective of this study was to investigate connec-tions between single nucleotide polymorphisms (SNP) of Estrogen receptor-α (ESR-α) with reproduction performance (calving interval, service per conception, and 305 d milk yield) in Hanwoo and Holstein dairy cattle. Hanwoo and Holstein blood samples were collected from 183 and 124 dam of breeding farms and DNA was extracted. Primer design was based on NCBI GenBank (Accession No. AY340579). The PCR-RFLP method with Bgl I was used to ge-notype the cattle. The result showed two variants of the ESR-α gene. The Bgl I cut the 492 bp amplification pro- duct into 322 bp and 170 bp fragments for allele G, while allele A remained uncut, resulting in two restriction frag-ments for homozygote G/G and three fragments for heterozygote A/G. We found two of different genotypes in the-se breeds, A/G and G/G. In Hanwoo, the A/G genotype frequency was 0.13, and G/G was 0.87. The CI of A/G was 382.18±10.03 days, and G/G was 381.69±5.22 days. The SPC of A/G was 1.62±0.16, and G/G was 1.32±0.04. While CI showed no significance difference, SPC exhibited significant difference (p<0.05). In Holstein cattle, the frequency of genotype A/G was 0.02 and G/G was 0.98. The 305 days milk yield of A/G was 7,253.00±936.00 kg and of G/G was 8,747.51±204.88 kg, showing no significant difference.
본 시험은 무항생제 사육방식이 한우육내 코티졸과 지방산 함량에 미치는 영향을 구명하고자 실시하였다. 무항생제 및 관행 축산으로 생산된 한우육 1등급 등심(M. longissimus)부위를 3가지의 브랜드별로 각각 3회씩 구매하여 시료로 공시하였고, 분석한 결과는 다음과 같다. 한우육 등심내 코티졸 함량은 무항생제 처리구에서 관행사육 처리구 대비 유의적(p=0.0176)으로 낮게 나타났으며, 포화지방산 및 불포화지방산 함량에서는 유의적(p>0.05)인 차이를 나타내지는 않았으나, 관행사육 처리구에서는 포화지방산이, 무항생제 처리구에서는 불포화지방산이 각각 높은 경향으로 분석되었다. 또한, 불포화지방산 중 n-3과 n-9 지방산을 제외한 n-6 지방산(linoleic acid, γ-lino-lenic acid)에서 유의적(p<0.01)인 차이를 보였다. 또한, 한우육내 코티졸 농도와 n-6 지방산 농도간의 상관관계를 분석한 결과, 특히 linoleic acid와 유의적(p=0.0140)으로 부(—)의 상관관계가 성립되었다.결과적으로, 무항생제 인증으로 사육된 한우육은 관행적으로 사육된 한우육에서 보다 상대적으로 코티졸 함량이 낮았으며, 건강에 유익한 불포화지방산 함량이 다소 높은 경향으로 분석됨으로써 육질을 향상시키는 것으로 나타났다.