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1995 Fiscal Year Final Research Report Summary

Development of embryo transfer system of bovine embryos sexed with PCR

Research Project

Project/Area Number 05556049
Research Category

Grant-in-Aid for Developmental Scientific Research (B)

Allocation TypeSingle-year Grants
Research Field Applied animal science
Research InstitutionKYOTO UNIVERSITY

Principal Investigator

YAMADA Masayasu  Kyoto Univ., Faculty of Agri., Associate Professor, 助教授, 助教授 (10243073)

Co-Investigator(Kenkyū-buntansha) MINAMI Naojiro  Kyoto Univ., Faculty of Agri., Researcher, 農学部, 助手 (30212236)
HOSOI Yoshihiko  Kinki Univ., Res.Inst.of Bio.-Oriented Sci.and Tech., Instructor, 生物理工学部, 講師 (70192739)
IRITANI Akira  Kinki Univ., Res.Inst.of Bio.-Oriented Sci.and Tech., Professor, 生物理工学部, 教授 (80026385)
GOTO Kazufumi  Kagoshima Univ., Faculty of Agri., Associate Professor, 農学部, 助教授 (30162142)
SUZUKI Tatsuyuki  Yamaguchi Univ., Faculty of Agri., Professor, 農学部, 教授 (00216409)
Project Period (FY) 1993 – 1995
KeywordsEmbryo transfer / Embryo sexing / Bovine SRY cDNA / PCR method / Cryopreservation / IVM / IVF bovine embryos
Research Abstract

Our objective was to test the practical use of direct transfer of frozen-thawed bovine embryos sexed with PCR method using bovine Y chromosome specific DNA probes. First, to develop bovine Y-specific DNA probes, cloning of bovine sex determining region of Y gene (bSRY), which is supposed to induce male differentiation of gonad during bovine embryogenesis was designed. As a result, using cloning techniques, such as RT-PCR with human SRY sequence as primers and total RNA extracted from bovine testes, and 5'RACE,two different sequences of bSRYcDNA were determined, but both had the evolutionary-conserved HMG box sequence. Therefore, the difference of the sequence may be attributed to the circular structure of bSRY mRNA,as has been described in mouse Sry. It was moreover found that the primers derived from bSRYcDNA were useful for sexing of bovine embryos with PCR method. Next, the transfer of frozen-thawed bovine embryos sexed with PCR method was directed. When the embryos biopsied were frozen using a freezing medium containing PVP as a cryoprotectant, the survival rate of embryos after thawing was very high and after transfer of those embryos, we could produce efficiently youngs with predicted sex.

  • Research Products

    (44 results)

All Other

All Publications (44 results)

  • [Publications] Utsumi, K.: "Embryo sexing by male specific antibody and by PCR using male specific (SRY) primer" Mol. Reprod. Dev.36. 238-241 (1993)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Utsumi, K.: "Embryo sex selection by a rat male-specific antibody and the cytogenetic and developmental confirmation in cattle embryos." Mol. Reprod. Dev.34. 25-32 (1993)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Takagi, M.: "Survival rate of frozen-thawed bovine IVF embryos in relation to exposure time using various cryoprotectants." Cryobiology. 30. 306-312 (1993)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Utsumi, K.: "Sexing of bovine embryos by PCR using bovine SRY primer" Theriogenology. 41. 323- (1994)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Minami, N.: "Nonspecies-specific effects of mouse oviducts on the development of boviane IVM/IVF embryos by a serum free co-culture" Theriogenology. 41. 1435-1445 (1994)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Suzuki, T.: "The influence of polyvinylpyrrolidone on freezing of bovine IVF blastocysts following biopsy." Cryobiology. 32. 505-510 (1995)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Nishikimi, A.: "Effects of anti H-Y mouse antisera on the blastocoel formation of rabbit embryos" Jpn. J. Embryo Transfer. 17. 88-93 (1995)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Iwata, H.: "Effect of taurine on bovine IVM-IVF embryo development co-cultured with cumulus cell" Jpn. J. Embryo Transfer. 17. 102-108 (1995)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Taniguchi, S.: "Hatching of bovine embryos clerived from in vitro fertilization in a cell-free, chemically defined medium." Jpn. J. Embryo Transfer. 17. 170-176 (1995)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Ookutsu, S.: "Direct transfer of bovine blastocysts derived from in vitro ferfilization after freezing and thawing by using ethylene glycol-trehalose as a cryoprotectant" Jpn. J. Embryo Transfer. 17. 177-182 (1995)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Saha, S.: "Viability of bovine blastocysts obtained after 7, 8 or 9 days of culture following vitrification and one-step rehydration" Theriogenology. 43. 311- (1995)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Tominaga, K.: "Influence of “Sorcoseryl" during culture on the sex dependent repair of bovine demi-embryos" Mol. Reprod. Dev.(in press).

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Utsumi, K.: "Sex determination by male specific antibody and by DNA amplification of mammalian embryos." J. Assist. Reprod. Genet.10. 62- (1993)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Takagi, M.: "Relationship between progesterone and estradiol contents of follicalar fluid and the merphological appearance of granubsa cells of follicles coexisting with corpola lutea in bovine ovanes" Theriogenology. 40. 135-147 (1993)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Kato, H.: "Effects of mouse oviducts on the development of bovine IVM/IVF embryos in a serum free medium." Theriogenology. 41. 224- (1994)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Takagi, M.: "Viability of inner cell mass of frozen-thawed IVM/IVF bovine embryos using various cryoproteetants." Theriogenology. 41. 310- (1994)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Goto, H.: "Sexing of bovine embryos by PCR amplification of biopsied trophoblast cell ant its clinical applification" Jpn. J. Embryo Transfer. 17. 19-26 (1995)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Kim, K. S.: "Effects of bovine follicular fluid on in vitro maturation of bovine oocytes." Jpn. J. Embryo Transfer. 17. 1-5 (1995)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Saha, H.: "Vitrification of bovine in vitro matured and pronuclear oocytes with different vitrification solutions" Theriogenology. 45. 179- (1996)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Fujitani, Y.: "Effects of oxygen tension, free radicals and hypotaurine on development of in vitro-derived bovine embryos." Theriogenology. 45. 210- (1996)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Kim, K. S.: "Functional role of cumulus cells during maturation in development of in vitro matured and fertilized bovine oocytes" Theriogenology. 45. 278- (1996)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Tominaga, K.: "Sex of IVM, IVF and IVC blastocysts produced discrimirately every donor cows" J. Reprod. Dev.(in press).

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Utsumi, K.: "Enbryo sexing by male specific antibody and by PCR using male specific (SRY) primer." Mol.Reprod.Dev.36. 238-241 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Utsumi, K.: "Embryo sex selection by a rat male-specific antibody and the cytogenetic and developmental confirmation in cattle embryos." Mol.Reprod.Dev.34. 25-32 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Utsumi, K.: "Sex determination by male specific antibody and by DNA amplification of mammalian embryos." J.Assist.Reprod.Genet.10. 62 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Takagi, M.: "Survival rate of frozen-thawed bovine IVF embryos in relation to exposure time using various cryoprotectants." Cryobiology. 30. 306-312 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Takagi, M.: "Relationship between progesterone and estradiol contents of follicular fluid and the morphological appearance of granulosa cells of follicles coexisting with corpola lutea in bovine ovaries." Theriogenology. 40. 135-147 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Utsumi, K.: "Sexing of bovine embryos by PCR using bovine SRY primer" Theriogenology. 41. 323 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Kato, H.: "Effect of mouse oviducts on the development of bovine IVM/IVF embryos in a serum free medium." Theriogenology. 41. 224 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Minami, N.: "Nonspecies-specific effects of mouse oviducts on the development of bovine IVM/IVF embryos by a serum free co-culture." Theriogenology. 41. 1435-1445 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Takagi, M.: "Viability of inner cell mass of frozen-thawed IVM/IVF bovine ebryos using various cryoprotectants." Theriogenology. 41. 310 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Goto, H.: "Sexing of bovine embryos by PCR amplification of biopsied trophoblast cell and its clinicalapplication." Jpn.J.Embryo Transfer. 17. 19-26 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Kim.K.S.: "Effects of bovine follicular fluid on in vitro maturation of bovine oocytes." Jpn.J.Embryo Transfer. 17. 1-5 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Nishikimi, A.: "Effects of anti H-Y mouse antisera on the bkastocoel formation of rabbit embryos." Jpn.J.Embryo Transfer. 17. 88-93 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Iwata, H.: "Effect of taurine on bovine IVM-IVF embryo development co-cultured with cumulus cell." Jpn.J.Embryo Transfer. 17. 102-108 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Taniguchi, S.: "Hatching of bovine embryos derived from in vitro fertilization in a cell-free, chemically defined medium." Jpn.J.Embryo Transfer. 17. 170-176 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Ookutsu, S.: "Direct transfer of bovine blastocysts derived from in vitro fertilization after freezing and thawing by using ethylene glycol trehalose as a cryoprotectant." Jpn.J.Embryo Transfer. 17. 177-182 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Saha, S.: "Viability of bovine blastocysts obtained after 7,8 or 9 days of culture following vitrificatin and one-step rehydration." Theriogenology. 43. 311 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Suzuki, T.: "The influence of polyvinylpyrrolidone on freezing of bovine IVF blastocysts following biopsy." Cryobiology. 32. 505-510 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Tominaga, K.: "Influence of "Solcoseryl" during culture on the sex-dependent repair of bovine demi-embryos." Mol.Reprod.Dev.(in press).

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Tominaga, K.: "Sex of IVM,IVF and IVC blastocysts produced discriminately every donor cows." J.Reprod.Dev.(in press).

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Saha, S.: "Vitrification of bovine in vitro matured and pronuclear oocytes with different vitrification solutions." Theriogenology. 45. 179 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Fujitani, Y.: "Effects of oxygen tension, free radicals and hypotaurine on development of in vitro-derived bovine embryos." Theriogenology. 45. 210 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Kim, K.S.: "Functional role of cumulus cells during maturation in development of in vitro matured and fertilized bovine oocytes." Theriogenology. 45. 278 (1996)

    • Description
      「研究成果報告書概要(欧文)」より

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Published: 1997-03-04  

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