• 제목/요약/키워드: oviduct

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In Vitro Maturation of Round Spermatids Using Porcine Oviduct Epithelial Cell Monolayer Condition Medium

  • Jabed Md. Anower;Kamal Tania;Lee Seung-Min;Kim Byung Ki
    • Reproductive and Developmental Biology
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    • v.29 no.4
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    • pp.241-245
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    • 2005
  • 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.

Purification of Embryogenesis Stimulating Activity from Bovine Oviduct Epithelial Cell Conditioned Medium and Its Effect on the Development of Bovine Embryos (소 난관상피세포 배양액 유래 배발육촉진인자의 정제와 소 수정란 체외배양시 발육에 미치는 영향)

  • 김현일;노상호;박종임;신태영;이병천;황우석
    • Journal of Embryo Transfer
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    • v.13 no.1
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    • pp.43-52
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    • 1998
  • The embryogenesis stimulating activity(ESA) had been shown in co-culture of embryos with bovine oviduct epithelial cell(BOEC) and culture in BOEG-conditioned medium. The present study was undertaken to purify and quantify the embryotropic proteins and to determine the optimum concentration of the embryotropic protein for the proper development of embryos. In BOEC-conditioned medium, five major bands of proteins were detected(66, 53, 40, 32 and 24 kDa) by SDS-PAGE. From these proteins, 288pg of protein that had a 32kDa molecular weight was purified by gel filtration column and perfusion chromatography ion-exchange column. When purified protein was supplemented to the in vitro culture media at various concentrations in protein-free media, 2.5$\mu$g /ml supplement group showed significantly higher rates of embryo development into morula /blastocyst stages than other groups(p<0.05). In conclusion, we purified 32kDa protein from BOEC-conditioned medium and this protein showed optimum embryogenesis stimulating effect at 2.5$\mu$g /ml.

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Studies on the Fertilized Eggs in the Internal Reproductive Organ of the Silkworm Female Moth (Bombyx mori L.) (자아의 내부 생식기내 수정란에 관한 연구)

  • 윤종관;사기언
    • Journal of Sericultural and Entomological Science
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    • v.13 no.2
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    • pp.119-122
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    • 1971
  • The following results were obtained through the experiments and study of fertility for silkworm eggs in vagina, oviduct, and ovarial base. The sample eggs were collected after leaving silkworm moth for one week from the date of treatment was given to oviporm orifice of the silkworm moth by heated needle in order to discontinue laying eggs. Those sample silkworm moths were divided into two groups. The first group was under the condition of immediately after the couplation and the second group was under the condition of after the laying eggs in some extend, and both of them were completed couplation. The sample eggs were screened in order to confirm the degree of color which normally be observed as very characteristic during hibernating. 1. About 5% of silkworm out of the total sample were identified as having 1-2 eggs. (See table 1). But, the charasteristic colour of hibernating egg was not observed. The colour was not much different as that of eggs in ovarian duct. 2. Immediately after the couplation, the eggs were not found in vagina under the condition of immediately after the couplation. 3. About 10% of the sample under the condition of after laying eggs in some extend were found having eggs in vagina or oviduct. 4. About 2% of the sample silkworm moth under the condition of immediately after the couplation were found having I fertilized egg in oviduct. 5. No fertilized egg was found in ovarian tube. It seemed that sperm did not enter ovarian tube. 6. Almost the entire eggs were found in ovarian tube under the condition of immediately after the couplation. But, eggs under the condition of after laying eggs in some extend were found around oviduct and vagina due to the continuous movement of silkworm eggs towards oviporus orifice. 7. Sperm is mainly found around seminal receptacle and vagina, and sperm goes into micropyle together with silkworm eggs which were moving towards oviduct.

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Expression of Neurotrophin 4 and Its Receptor Tyrosine Kinase B in Reproductive Tissues during the Follicular and Luteal Phases in Cows

  • Sun, Yongfeng;Li, Chunjin;Sun, Yanling;Chen, Lu;Liu, Zhuo;Ma, Yonghe;Wang, Chunqiang;Zhang, Wei;Zhou, Xu
    • Asian-Australasian Journal of Animal Sciences
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    • v.24 no.3
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    • pp.336-343
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    • 2011
  • The neurotrophins, required for the survival and differentiation of the nervous system, are known to be important for the development of the reproductive tissues. However, the signals initiating the growth of follicles, gamete development, and transport and the development of zygote in the reproductive system of cows remain ambiguous. The purpose of the present study was to identify the transcripts and proteins of Neurotrophin 4 (NT4) and its receptor tyrosine kinase B (TrkB) in bovine reproductive tissues. The transcripts and immunoreactivity of NT4 and TrkB proteins were detected by reverse transcription polymerase chain reaction and western blot analysis. Using immunohistochemistry, the specific immunoreactivity of NT4 and TrkB were detected in the oocytes of primordial follicles and in the growing primary follicles. The NT4 and TrkB immunoreactivity was predominantly observed in granulosa cells, cumulus granulosa cells, cumulus oocyte complexes, theca cells of mature follicles, as well as in the oviduct epithelial cells, uterine gland cell, and epithelium cells of the uterus during the follicular and luteal phases in cows. Expressions of NT4 and TrkB mRNAs were not significantly different among the ovary, oviduct, and uterus of the follicular phase. For the luteal phase, the expression of NT4 mRNA in the ovary was significantly higher than that in the oviduct and uterus, and the expression of TrkB mRNA in the oviduct was significantly higher than that in the ovary and uterus, as determined by fluorescence quantitative reverse transcription polymerase chain reaction. The expression of NT4 mRNA was significantly higher than that of TrkB mRNA in the ovary and uterus, whereas NT4 mRNA expression was lower than that of TrkB mRNA in the oviduct during the luteal phase. The present study hypothesizes that NT4 participates in the regulation of both gonads and extra-gonadal reproductive tissues in cows.

Studies on the culture of bovine embryos using synthetic oviduct fluid(SOF) (Synthetic oviduct fluid(SOF)를 이용한 소 수정란의 배양에 관한 연구)

  • Roh, Sang-ho;Hwang, Woo-suk;Jo, Choong-ho
    • Korean Journal of Veterinary Research
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    • v.35 no.1
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    • pp.187-195
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    • 1995
  • The present study carried out to determine the developmental capacity of bovine oocytes matured in epidermal growth factor(EGF)-containing medium, the developmental competence of bovine embryos using synthetic oviduct fluid(SOF) and the effect of glucose on the development of bovine embryos. In experiment 1, oocytes, obtained from abattoir ovaries, were matured in EGF-containing medium for 24 hours, followed by exposure to Korean native cattle spermatozoa for 18 hours and cultured by utilizing co-culture system with bovine oviduct epithelial cells(BOEC) in TCM199. In experiment 2, early bovine embryos were cultured in SOF with or without BOEC and compared with those in TCM199 with BOEC. In experiment 3, bovine embryos were cultured in the presence or absence of glucose. Seven and ten days after in vitro fertilization, developmental competence of embryos were evaluated. The rate of cleavage was significantly(P<0.05) higher in EGF-containing maturation medium(70.0%) than in control(57.7%). The rates of development to morulae and blastocysts were 30.6% and 23.3% there was no significant difference between them. The rates of in vitro fertilized embryos to morulae and blastocysts cultured in SOF with BOEC(30.4%) and in TCM199 with BOEC(38.0%) were significantly(P<0.01) higher than cultured in SOF without BOEC(13.4%) at seven days after in vitro fertilization. The rates of embryos to blastocysts cultured in SOF with BOEC(29.4%) and in TCM199 with BOEC(35.9%) were significantly(P<0.05) higher than cultured in SOF without BOEC(13.4%) at ten days after in vitro fertilization. The rates of early embryos to morulae and blastocysts cultured in the presence or absence of glucose were 12.2% and 17.5% each other, there was no significant difference between them. The results show that bovine oocytes matured in the presence of EGF can cleave better, SOF with BOEC can replace serum containing complex media, TCM199 with BOEC in bovine embryo culture and glucose have little effect on the culture of early bovine embryos.

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Investigational Studies on Reproductive Failures of Slaughtered Cows (도살빈우의 번식장애사례 조사연구)

  • 이용빈;임경순
    • Korean Journal of Animal Reproduction
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    • v.6 no.1
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    • pp.19-30
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    • 1982
  • 1. The cows slaughtered at age of 3, 4, 6, 7, 8, and 9 years old were 1.5, 1.5, 15.0, 62.5 and 4.4% respectively. 2. The cows slaughtered at 351-450kg and more than 500kg were 60 and 28% respectively. 3. Best, very good, good and bad cows in nutritional condition were 1.6, 25.8, 62.9, and 9.7% respectively. Among the six cows which were bad nutrition, the two were with severe endometritis, the three were normal in genital function and one was on 70 days of pregnancy. 4. Holstein cows(55.2%) showed higher reproductive failure than the Korean cows(33.3%). 5. The slaughted ratio of the Korean cattle and Holstein cows was 36 and 64% respectively. 6. Pregnant cows were about 16% among the slaughtered one. 7. Reproductive failures were composed of 46% in uterus, 32% in ovaries, 8% in udder, 6% in oviduct, 4% in cervix of uterine, 2% in vagina and 2% inmummified fetus. 8. Forty six percentages of uterine diseases were as follows; horn, 13%, body of uterus, 32% and ovary diseases were 32%, that is, 12% of ovary atrophy, 8% of ovarycyst and 6% of lutealcyst. 9. The cows of reproductive failures were commonly infected with 1.6 kinds of diseases. 10. According to classification, six type of ovaries were as follows; normal, 58%, ovary-cyst, 11%, luteum cyst, 4%, coexistence of follicles and corpus luteum, 16%, weak function of ovaries, 10% and ovarian atrophy, 1%. 11. Major axis, minor axis and thickness of right ovary were larger than those of left one both in Korean cattle and Holstein cows. Holstein cow had generally larger size of ovary than these of the Korean cattle.. 12. The left and right oviducts showed no difference in length, but Holstein had longer oviduct than Korean cow. 13. There was no difference in the length of uterine horn between right and left in the Korean cows, but the right was longer than the left in Holstein cows. 14. Holstein had longer horn and body of uterine than the Korean cows. 15. The weight of right ovary was heavier than that of left in both breeds, but there was no differences in weight of left ovary between two breeds and right ovary of Holstein breed was heavier than that of the Korean cow. 16. The weight of right oviduct and uterine born was heavier than that of the left, and Holstein had heavier oviducts and uterine horns than the Korean cows. 17. Holstein had heavier uterine body and cervix of uterine than the Korean cows. 18. The length of reproductive systems of Korean cow is as follows; Major and minor diameter and thickness ofovary are 3.6${\pm}$0.7, 2.3${\pm}$0.4 and 1.6${\pm}$1.4 cm in left and 3.7${\pm}$0.6, 2.5${\pm}$0.5 and 1.8${\pm}$0.5 cm in right. Oviduct is 28.4${\pm}$3.1 cm in left and 27.8${\pm}$3.3 cm in right. Uterine horn is 27.4${\pm}$4.5 cm in left and 27.7${\pm}$4.9 cm in right. Uterine body and cervix are 3.4${\pm}$1.1 and 6.5${\pm}$1.7 cm. 19. The length of female reproductive systems ofHolstein cow is as follows; Major and minor diameter and thickness of ovary are 3.9${\pm}$1.3, 2.3${\pm}$0.5, and 1.5${\pm}$0.6 cm in left and 4.0${\pm}$0.8, 2.8${\pm}$0.6 and 1.8${\pm}$0.6 cm in right. Oviduct is 29.4${\pm}$4.2 cm in left and 29.3${\pm}$4.1 cm in right. Uterine horn is 30.2${\pm}$7.4 cm in left and 32.6${\pm}$8.4 cm in right. Uterine body and cervix are 4.5${\pm}$2.5 and 7.8${\pm}$2.9 cm. 20. The weight of reproductive systems of Korean cow is as follows; Ovary is 8.4${\pm}$4.1 g in left and 9.3${\pm}$3.6g in right. Oviduct is 1.5${\pm}$0.5 g in left and 1.6${\pm}$0.5 g in right. Uterine horn is 109${\pm}$27 g left and 118${\pm}$32 g in right. Uterine body and cervix are 30.4${\pm}$14.1 and 76.7${\pm}$38.4g. 21. The weight of reproductive systems of Holstein cow is as follows; Ovary is 8.2${\pm}$3.1 g in left and 12.5${\pm}$5.6 g in right. Oviduct is 1.7${\pm}$0.6 g in left and 1.9${\pm}$0.9 g in right. Uterine horn is 199${\pm}$14.2 g in left and 221${\pm}$111.2g in right. Uterine body and cervix are 58.2${\pm}$46.5 and 126.7${\pm}$103.3 g.

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Development Capacity of Bovine Oocytes Matured and Fertilized In Vitro (체외성숙 및 수정된 우 난포란의 발생능)

  • 서태광;박수봉;박항균
    • Korean Journal of Animal Reproduction
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    • v.14 no.1
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    • pp.17-22
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    • 1990
  • The present study was carried to investigate the development potential of bovine follicular oocytes matured and fertilized in vitro. Bovine oocytes matured in vitro were fertilized, cultured, and transfered to the rabbit oviduct for in vitro culture to evaluate the development potential. The rates of in vitro maturation, fertilization and polyspermy were 96%(355/369), 90%(320/355) and 15%(17/320), respectively. The percentage of oocytes cleaved after culture for 48 hours, to 2 cell, 3~4 cell, 6~8 cell stage were 17%(60/356), 21%(75/356) and 19%(67/356), respectively and overall cleavage rate was 57%(202/356). The rate of recovered embryos after 5 days in rabbit oviduct was 56%(80/142), and 21%(17/80) of recovered embryos developed over morula stage.

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Expression and Function of CTNNB1 in the Development of Avian Reproductive System

  • Bae, Seung-Min;Song, Gwonhwa
    • Reproductive and Developmental Biology
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    • v.38 no.1
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    • pp.35-40
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    • 2014
  • Beta-catenin (CTNNB1, catenin (cadherin-associated protein), beta 1) is involved in various biological processes, including embryogenesis, tumorigenesis, angiogenesis and progression of metastasis. CTNNB1, as a multifunctional and oncogenic protein, has important roles in adhesion between Sertoli cells through an N-cadherin-dependent manner and in various cancer types through its over-activation. In addition, CTNNB1 can interact with estrogen/estrogen receptor alpha complex, which regulates the transcription of WNT (wingless-type MMTV integration site family)/CTNNB1 target genes. Recently, we investigated the functional roles and expression pattern of CTNNB1 during the morphological changes of embryonic gonads of chickens and the estrogen-dependent regulation of CTNNB1 in oviduct development and potential functions as a biomarker of CTNNB1 in human epithelial ovarian cancer using the chicken as a biological research model. Therefore, in this review, we provide a new insight of potential role of CTNNB1 in the development of the female reproductive tract during early embryogenesis and ovarian carcinogenesis of laying hen models.

Gene Gun-Mediated Human Erythropoietin Gene Expression in Primary Cultured Oviduct Cells from Laying Hens

  • Ochiai, H.;Park, H.M.;Sasaki, R.;Okumura, J.;Muramatsu, T.
    • Asian-Australasian Journal of Animal Sciences
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    • v.12 no.1
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    • pp.9-14
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    • 1999
  • Factors affecting gene gun-mediated expression of the human erythropoietin gene were investigated in primary cultured oviduct cells from laying hens. The human erythropoietin gene was transfected by a gene gun method at $1.25{\mu}g$ per dish, and cultured in a synthetic serum-free medium for 72 hrs. The concentration of human erythropoietin mRNA was determined by RNA : RNA solution hybridization. In experiment 1, the effect of changing the shooting pressure of DNA-coated microparticles with nitrogen gas was tested at 20 and $60kgf/cm^2$. The results showed that the erythropoietin mRNA concentration was significantly higher at 60 than $20kgf/cm^2$. In experiment 2, the effects of supplementing the medium with fetal calf serum at 10%, and raising the shooting pressure from 60 to $80kgf/cm^2$ on the cell number and erythropoietin gene expression were examined. Although supplementation with fetal calf serum significantly increased the cell numbes compared with no supplemented controls (p < 0.05), erythropoietin mRNA concentration per $10^3$ cells was not affected. Raising the shooting pressure from 60 to $80kgf/cm^2$ did not affect either of the parameters, In experiment 3, the effect of supplementing ascorbate 2-phosphate at 0.5 mM was tested. The results indicated that the ascorbate supplementation significantly increased the cell number (p < 0.05), and tended to increase erythropoietin mRNA concentration (p < 0.1). Thus, for human erythropoietin gene expression by using the gene gun method, shooting pressure with nitrogen gas should be sufficient at $60kgf/cm^2$ and supplementation with ascorbate phosphate would be useful to enhance not only the cell proliferation but also erythropoietin gene expression.