간행물
Reproductive & developmental biology
- 발행기관 한국동물번식학회
- 자료유형 학술지
- 간기 계간
- ISSN 1738-2432 (Print)2288-0151 (Online)
- 수록기간 1977 ~ 2018
- 주제분류 농수해양 > 축산학 농수해양 분류의 다른 간행물
- 십진분류KDC 527DDC 636
권호리스트/논문검색
Volume 29 No 4 (2005년 12월) 10건
1.
2005.12
구독 인증기관 무료, 개인회원 유료
In the present study, we investigated the effects of genotypes on in vitro maturation and fertilization in porcine fresh/frozen-thawed oocytes. The porcine cumulus-oocyte complexes (COCs) were divided into four groups according to whether they were: (1) in vitro matured; (2) cryopreserved and in vitro matured; (3) in vitro fertilized and (4) cryopreserved, and in vitro fertilized. Maturation of porcine COCs was accomplished by incubation in NCSU23 medium. Immature oocytes were cryopreserved by Open Pulled Straws (OPS) method according to Vajta et al., (1998). Oocytes stained by Acetic-Orcein method were observed under the microscope. DNA extracted from the ovaries was analyzed by RAPD (random amplified polymorphic DNA) and SSCP (single strand conformational polymorphisrrt) method. The rates of oocytes maturation and fertilization were significantly high in AA genotype. The results indicated that in vitro maturation and fertilization in porcine fresh/frozen-thawed oocytes may be affected by genotypes in pigs.
4,000원
2.
2005.12
구독 인증기관 무료, 개인회원 유료
We used nine decamer primers to generate DNA fragment sizes ranging from 100 bp to 1,600 bp from two bullhead (Pseudobagrus fulvidraco) populations of Dangjin in Korea. 376 fragments were identified in the cultured bullhead population, and 454 in the population of wild bullhead from Dangjin: 287 specific fragments (76.3%) in the cultured bullhead population and 207 (45.6%) in the wild bullhead population. On average, a decamer primer was used to generate 34.2 amplified products in a cultured bullhead. A RAPD primer was used to generate an average of 3.1 amplified bands per sample, ranging between 2.5 and 6.0 fragments in this population. Nine primers also generated 24 polymorphic fragments (24/376 fragment, 6.4%) in the cultured bullhead population, and 24 (24/454 fragments, 5.2%) in the wild bullhead population. The OPA-16 primer, notably, produced which 11 out of 11 bands (100%) were monomorphic in the wild bullhead population. 110 intra-population-specific fragments, with an average of 12.2 per primer, were observed in the cultured bullhead population. 99 fragments, with an average of 11.0 per primer, were identified in the wild bullhead. Especially, 55 inter-population-common fragments, with an average of 6.1 per primer, were observed in the two bullhead populations. The bandsharing value (BS value) of individuals within the wild bullhead population was substantially higher than was determined in the cultured bullhead population. The average bandsharing value was 0.596±0.010 within the cultured bullhead population,. and 0.657±0.010 within the wild bullhead population. The dendrogram obtained with the nine primers indicates two genetic clusters, designated cluster 1(CULTURED 01~CULTURED 11), and cluster 2(WILD 12~WILD 22). Ultimately, the longest genetic distance displaying significant molecular differences was determined to exist between individuals in the two bullhead populations, namely between individuals WILD no. 19 of the wild bullhead population and CULTURED no. 03 of the cultured bullhead population (genetic distance = 0.714). RAPD-PCR allowed us to detect the existence of population discrimination and genetic variation in Korean population of bullhead. This finding indicates that this method constitutes a suitable tool for DNA comparison, both within and between individuals, populations, species, and genera.
4,000원
3.
2005.12
구독 인증기관 무료, 개인회원 유료
In vitro maturation of porcine immature cumulus-enclosed oocytes can be enhanced by co-incubation with spermatozoa even before fertilization. The aim of this study was to determine whether the addition of spermatozoa into the culture medium can stimulate the meiosis resumption of porcine cumulus-enclosed oocytes arrested at germinal vesicle (GV). Cumulus-enclosed oocytes (CEOs) were collected from follicles of 3 to 5mm diameter. Porcine CEOs were cultured in tissue culture medium containing various meiosis inhibitors and spermatozoa. Oocytes were examined for evidence of GV and GV breakdown after 24 h culture. After 24 h culture 43.8% of oocytes cultured in only TCM 199 remained at GV stage whereas 56.2% of oocytes were able to resume meiosis. When porcine CEOs were cultured in the medium with meiosis inhibitor such as, dibutyryl cAMP (dbcAMP) and forskolin (Fo), more than 90% of oocytes were not able to resume meiosis. However, co-culture of porcine CEOs with spermatozoa was able to overcome the inhibitory effect of dbcAMP and Fo. Irrespective of the presence of 3-isobutyl-1-methylxanthine (IBMX), no difference was observed in the proportion of oocyte reached germinal vesicle breakdown (GVBD). The present study suggests that dbcAMP and Fo prevent the spontaneous maturation of competent oocyte in culture after isolation from follicles and that mammalian spermatozoa contain a substance(s) that improves meiosis resumption in vitro of porcine cumulus-enclosed oocytes.
4,000원
4.
2005.12
구독 인증기관 무료, 개인회원 유료
Methods for activation of reconstructed oocytes were examined for the production of nuclear transfer (NT) rat embryos using fetal neural stem cells as donor. Neural stem cells were isolated from Day 14.5 rat fetuses, and the oocytes for recipient cytoplasm were recovered from 4-week old Sprague Dawley rats. After enucleation and nuclear injection, the reconstructed oocytes were immediately exposed to activation medium consisting of 10 mM SrCl₂ for 4 h (immediate activation after injection; IAI), or cultured in vitro for 2~3 h before activation treatment (injection before activation; IBA). Pre-activated oocytes were also used for NT to test reprogramming potential of artificially activated oocytes. The oocytes were grouped as IIA (immediate injection after activation) and ABI (activation 2~3 h before injection). Following NT, the oocytes were cultured in vitro. Development of the NT embryos was monitored at 44 and 119 h after activation. The embryos in groups IAI, mA, and IIA were cleaved to the 2-cell stage at the rates of 36.6%(15/41), 39.5% (17/43) and 46.3% (25/54), respectively. However, in the ABI group, only one embryo (1.8%, 1/55) was cleaved after activation. After in vitro culture, two NT embryos from IAI group had developed to the morula stage (4.9%, 2/41). However, no morula or blastocyst was obtained in the other groups. These results suggest that immediate activation after injection (IAI) method may be used for the production of rat somatic cell NT embryos.
4,000원
5.
2005.12
구독 인증기관 무료, 개인회원 유료
This study was conducted to test whether the transgenic cattle pass the transgene to their progeny through germ cells, and whether the transgene is expressed in the mammary gland of ransgenic cows. Two male ransgenic calves were born from IVF-derived embryos injected with bovine β-casein/human lactoferrin fusion gene and then grew up to be reproducible. Semen was collected from a transgenic bull after 18 mon of age and then frozen. Bovine oocytes matured in vitro were fertilized with spermatozoa of the transgenic bull and cultured in 50㎕ drops of CRlaa medium supplemented with 3 mg/mL BSA. After 48 h of culture, cleaved embryos were determined for the presence of transgenes by DNA polymerase chain reaction (PCR). Proportion of transgene positives among bovine embryos fertilized with sperm of the transgenic bull was 20.9% (28/134). One of transgenic bulls did not produce transgenic sperm. Out of 34 calves produced from recipient heifers inseminated with semen of the other bull, 3 (8.8%) were transgenic animals (2 females and 1 male). Thus, one transgenic bull showed a low transmission frequency below Mendelian levels in both the IVF-derived embryos and his progeny. It was demonstrated by Southern blot that copy numbers of the transgene in the transgenic progeny enhanced about 1.8 times as compared to those of the founder bull The results demonstrate that the transgenic bull carrying human lactoferrin gene could pass his transgene to the progeny through germ cells, although he is a germ-line mosaic.
4,000원
6.
2005.12
구독 인증기관 무료, 개인회원 유료
Porcine oviduct epithelial cells (POEC) are widely used in co-culture experiments to improve early embryonic development, in vitro fertilization in embryo transfer programs for domestic animals and in vitro maturation of immature germ cells. POEC were mechanically isolated and cultured in tissue culture medium 199. Cells grew continuously, and confluent monolayers were formed after 7 days. After forming confluent monolayer of epithelial cells, supernatant was collected as the condition medium for maturing round spermatids in vitro. Round spermatids were also separated mechanically and cultured in the POEC condition medium. In this study we observed that 20% of round spermatid cultured were matured into elongating spermatid after 24 h, and about 10% of round spermatid cultured showed complete elongation (elongated spermatid) within 24~48 h of in vitro culture. No further development was observed within 50~72 h and transformed cells lost their viability after 72 h. These preliminary findings suggest that the condition medium from POEC may be possible to overcome the round spermatid block by improving the milieu of culture system.
4,000원
7.
2005.12
구독 인증기관 무료, 개인회원 유료
The purpose of this study was to assess sperm quality during in vitro storage of miniature-pig semen in order to determine which extender should be used and how extender can be diluted for in vitro storage of miniature-pig semen. Freshly ejaculated miniature-pig's semen was diluted with same volumes of Beltsville Thawing Solution (BTS), Androhep, Modena, Mulberry III and modified-Modena extenders. Sperm quality was evaluated by examining viability, motility, abnormality, acrosome intactness, intensity and capacitation status by chlorotetracycline (CTC) staining. Sperm motility decreased with storage period prolonged and differences among BTS, Androhep, Modena and Mulberry III were apparent On Day 1, approximately 80% of the sperm were motile, but motility decreased to 40% at Day 7. During the 7 days of storage, sperm survival in modified-Modena B extender was higher than another extenders. However, it was not differ significantly among other extenders. The percentage of F and B patterns were not differ significantly among the extenders. However, F pattern in modified-Modena B extender was slightly higher until 3 days of storage than that of Modena extender, modified-Modena A extender and modified-Modena C extender. The percentage of AR patterns in modified-Modena B extender was slightly lower, but did not differ significantly among other extenders. The results of present study suggest that modified-Modena B was effective as new extender for in vitro storage of miniature-pig semen.
4,000원
8.
2005.12
구독 인증기관 무료, 개인회원 유료
This study was carried out to evaluate the effects of washing medium, breed and washing temperature of fresh and frozen-thawed boar sperm on mitochondrial activity and membrane integrity by flow cytometry. More than 80% of fresh sperm washed with mTLP-PVA medium at 20℃ exhibited an intact membrane and a functional mitochondrion. With frozen-thawed samples, a large number of sperm showed both damaged membrane (36.4~46.9%) and nonfunctional mitochondrion (55.1~71.1%) in the mTLP-PVA and BTS washing media at 20℃. There were no breed effects of fresh and frozen-thawed sperm on mitochondrial activity and membrane integrity. The percentages of damaged membrane of fresh and frozen sperm, respectively, were higher at 4℃ washing temperature than at 20℃ washing temperature in the mTLP-PVA medium. We found that washing medium and washing temperature of fresh and frozen-thawed boar sperm were important for the analyses of mitochondrial activity and membrane integrity by flow cytometry.
4,000원
9.
2005.12
구독 인증기관 무료, 개인회원 유료
The present study was conducted to investigate gelatinolytic activities in HAM and to determine whether there are any changes in gelatinolytic activity profiles when the cells are cultured in hepatogenic medium. Placenta was obtained during caesarean section of the volunteers, with informed consent. HAM were isolated from amniotic membrane using collagenase type A HAM were cultured in hepatogenic medium for 3 weeks and the conditioned media were obtained at day 7, 14 and 21. The zymographic pattern of gelatinolytic activity of the HAM did not undergo a change during passages. When the HAM were cultured in a fibronectin-coated dishes in a hepatogenic medium, there was no significant difference of the gelatinase pattern between before and after culture. However, when bFGF was added to the culture, a dramatic increase of 62kDa and 59kDa gelatinases was observed. Interestingly, when ITS instead of FN was present, HAM-conditioned medium also showed a similar increase of both gelatinases. Immunoblotting analysis demonstrated that both 62kDa and 59kDa gelatinases were the active form of MMP-2 resulting from the turnover of MMP-2 proform. Futher study will be necessary to determine the relationship between bFGF and active MMP-2 during hepatogenesis of HAM.
4,000원
Short Communication
10.
2005.12
구독 인증기관 무료, 개인회원 유료
The freemartinism is the most frequent form of intersexuality found in cattle, and females of heterosexual twins become sterile. With increase of twinning rates due to transfer of multiple embryos derived from in vitro fertilization, it is of great economic value to establish early diagnosis of freemartins to remove infertile individuals from breeding stock. In the present study polymerase chain reaction (PCR) of two different Y-chromosome specific segments (BRY.l and AMX/Y) was performed to identify freemartins from twins and less common single born freemartins in Korean Native Cattle (KNC). Two male-specific sequences were amplified in all heterosexual twins tested (n=5). In addition, Y-specific PCR products were detectable in one of the single born females (n=4) with visible genital abnormalities. These results suggest that the sensitivity of PCR-based assay may be sufficient to detect freemartinism in single born females as well as female partners of heterosexual twins in KNC.
3,000원
