Transient meiotic arrest maintained by DON (6-diazo-5-oxo-l-norleucine) enhances nuclear/cytoplasmic maturation of porcine oocytes
The developmental competence of in vitro-matured oocytes is still lower than that of the in vivo-matured oocytes due to precocious meiotic resumption and inappropriate cytoplasmic maturation. Although numerous efforts have been attempted to accomplish better in vitro maturation (IVM) condition, only...
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Published in | Reproduction (Cambridge, England) Vol. 158; no. 6; pp. 543 - 554 |
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Main Authors | , , , , , , , , , , , , , , , , |
Format | Journal Article |
Language | English |
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England
Bioscientifica Ltd
01.12.2019
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Abstract | The developmental competence of in vitro-matured oocytes is still lower than that of the in vivo-matured oocytes due to precocious meiotic resumption and inappropriate cytoplasmic maturation. Although numerous efforts have been attempted to accomplish better in vitro maturation (IVM) condition, only limited progress has been achieved. Thus, a current study was conducted to examine the effects of 6-diazo-5-oxo-l-norleucine (DON, an inhibitor of hyaluronan synthesis) during the first half period of IVM on nuclear/cytoplasmic maturation of porcine oocytes and subsequent embryonic development. Based on the observation of the nucleus pattern, metaphase II (MII) oocyte production rate in 1 µM DON group was significantly higher than other groups at 44 h of IVM. The 1 µM of DON was suggested to be optimal for porcine IVM and was therefore used for further investigation. Meiotic arrest effect of DON was maximal at 6 h of IVM, which was supported by the maintenance of significantly higher intra-oocyte cAMP level. In addition, increased pERK1/2 levels and clear rearrangement of cortical granules in membrane of MII oocytes matured with DON provided the evidence for balanced meiosis progression between nuclear and cytoplasmic maturation. Subsequently, DON significantly improved blastocyst formation rate, total cell numbers, and cellular survival in blastocysts after parthenogenetic activation, in vitro fertilization, and somatic cell nuclear transfer. Altogether, our results showed for the first time that 1 µM DON can be used to increase the yield of developmentally competent MII oocytes by synchronizing nuclear/cytoplasmic maturation, and it subsequently improves embryo developmental competence. |
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AbstractList | The developmental competence of in vitro-matured oocytes is still lower than that of the in vivo-matured oocytes due to precocious meiotic resumption and inappropriate cytoplasmic maturation. Although numerous efforts have been attempted to accomplish better in vitro maturation (IVM) condition, only limited progress has been achieved. Thus, a current study was conducted to examine the effects of 6-diazo-5-oxo-l-norleucine (DON, an inhibitor of hyaluronan synthesis) during the first half period of IVM on nuclear/cytoplasmic maturation of porcine oocytes and subsequent embryonic development. Based on the observation of the nucleus pattern, metaphase II (MII) oocyte production rate in 1 µM DON group was significantly higher than other groups at 44 h of IVM. The 1 µM of DON was suggested to be optimal for porcine IVM and was therefore used for further investigation. Meiotic arrest effect of DON was maximal at 6 h of IVM, which was supported by the maintenance of significantly higher intra-oocyte cAMP level. In addition, increased pERK1/2 levels and clear rearrangement of cortical granules in membrane of MII oocytes matured with DON provided the evidence for balanced meiosis progression between nuclear and cytoplasmic maturation. Subsequently, DON significantly improved blastocyst formation rate, total cell numbers, and cellular survival in blastocysts after parthenogenetic activation, in vitro fertilization, and somatic cell nuclear transfer. Altogether, our results showed for the first time that 1 µM DON can be used to increase the yield of developmentally competent MII oocytes by synchronizing nuclear/cytoplasmic maturation, and it subsequently improves embryo developmental competence. The developmental competence of in vitro -matured oocytes is still lower than that of the in vivo -matured oocytes due to precocious meiotic resumption and inappropriate cytoplasmic maturation. Although numerous efforts have been attempted to accomplish better in vitro maturation (IVM) condition, only limited progress has been achieved. Thus, a current study was conducted to examine the effects of 6-diazo-5-oxo-l-norleucine (DON, an inhibitor of hyaluronan synthesis) during the first half period of IVM on nuclear/cytoplasmic maturation of porcine oocytes and subsequent embryonic development. Based on the observation of the nucleus pattern, metaphase II (MII) oocyte production rate in 1 µM DON group was significantly higher than other groups at 44 h of IVM. The 1 µM of DON was suggested to be optimal for porcine IVM and was therefore used for further investigation. Meiotic arrest effect of DON was maximal at 6 h of IVM, which was supported by the maintenance of significantly higher intra-oocyte cAMP level. In addition, increased pERK1/2 levels and clear rearrangement of cortical granules in membrane of MII oocytes matured with DON provided the evidence for balanced meiosis progression between nuclear and cytoplasmic maturation. Subsequently, DON significantly improved blastocyst formation rate, total cell numbers, and cellular survival in blastocysts after parthenogenetic activation, in vitro fertilization, and somatic cell nuclear transfer. Altogether, our results showed for the first time that 1 µM DON can be used to increase the yield of developmentally competent MII oocytes by synchronizing nuclear/cytoplasmic maturation, and it subsequently improves embryo developmental competence. |
Author | Yoon, Seung-Bin Song, Bong-Seok Lee, Sang-Rae Choi, Seon-A Kim, Ji-Su Jeong, Pil-Soo Kang, Philyong Yang, Hae-Jun Park, Young-Ho Kim, Young-Hyun Sim, Bo-Woong Choo, Young-Kug Kim, Sun-Uk Huh, Jae-Won Jeong, Kang-Jin Koo, Deog-Bon Lee, Sanghoon |
AuthorAffiliation | Primate Resource Center, Korea Research Institute of Bioscience and Biotechnology, Jeollabuk-do, Republic of Korea National Primate Research Center, Korea Research Institute of Bioscience and Biotechnology, Chungcheongbuk-do, Republic of Korea Department of Biological Science, College of National Sciences, Wonkwang University, Jeollabuk-do, Republic of Korea Department of Functional Genomics, KRIBB School of Bioscience, Korea University of Science and Technology (UST), Daejeon, Republic of Korea Futuristic Animal Resource & Research Center, Korea Research Institute of Bioscience and Biotechnology, Chungcheongbuk-do, Republic of Korea Department of Biotechnology, Daegu University, Gyeongsangbuk-do, Republic of Korea |
AuthorAffiliation_xml | – name: Futuristic Animal Resource & Research Center, Korea Research Institute of Bioscience and Biotechnology, Chungcheongbuk-do, Republic of Korea – name: National Primate Research Center, Korea Research Institute of Bioscience and Biotechnology, Chungcheongbuk-do, Republic of Korea – name: Department of Biological Science, College of National Sciences, Wonkwang University, Jeollabuk-do, Republic of Korea – name: Department of Biotechnology, Daegu University, Gyeongsangbuk-do, Republic of Korea – name: Primate Resource Center, Korea Research Institute of Bioscience and Biotechnology, Jeollabuk-do, Republic of Korea – name: Department of Functional Genomics, KRIBB School of Bioscience, Korea University of Science and Technology (UST), Daejeon, Republic of Korea |
Author_xml | – sequence: 1 givenname: Hae-Jun surname: Yang fullname: Yang, Hae-Jun organization: Department of Biological Science, College of National Sciences, Wonkwang University, Jeollabuk-do, Republic of Korea – sequence: 2 givenname: Sanghoon surname: Lee fullname: Lee, Sanghoon organization: Futuristic Animal Resource & Research Center, Korea Research Institute of Bioscience and Biotechnology, Chungcheongbuk-do, Republic of Korea – sequence: 3 givenname: Bo-Woong surname: Sim fullname: Sim, Bo-Woong organization: National Primate Research Center, Korea Research Institute of Bioscience and Biotechnology, Chungcheongbuk-do, Republic of Korea – sequence: 4 givenname: Pil-Soo surname: Jeong fullname: Jeong, Pil-Soo organization: Department of Biotechnology, Daegu University, Gyeongsangbuk-do, Republic of Korea – sequence: 5 givenname: Seon-A surname: Choi fullname: Choi, Seon-A organization: National Primate Research Center, Korea Research Institute of Bioscience and Biotechnology, Chungcheongbuk-do, Republic of Korea – sequence: 6 givenname: Young-Ho surname: Park fullname: Park, Young-Ho organization: Department of Functional Genomics, KRIBB School of Bioscience, Korea University of Science and Technology (UST), Daejeon, Republic of Korea – sequence: 7 givenname: Bong-Seok surname: Song fullname: Song, Bong-Seok organization: Department of Functional Genomics, KRIBB School of Bioscience, Korea University of Science and Technology (UST), Daejeon, Republic of Korea – sequence: 8 givenname: Seung-Bin surname: Yoon fullname: Yoon, Seung-Bin organization: Department of Functional Genomics, KRIBB School of Bioscience, Korea University of Science and Technology (UST), Daejeon, Republic of Korea – sequence: 9 givenname: Philyong surname: Kang fullname: Kang, Philyong organization: National Primate Research Center, Korea Research Institute of Bioscience and Biotechnology, Chungcheongbuk-do, Republic of Korea – sequence: 10 givenname: Kang-Jin surname: Jeong fullname: Jeong, Kang-Jin organization: National Primate Research Center, Korea Research Institute of Bioscience and Biotechnology, Chungcheongbuk-do, Republic of Korea – sequence: 11 givenname: Young-Hyun surname: Kim fullname: Kim, Young-Hyun organization: Department of Functional Genomics, KRIBB School of Bioscience, Korea University of Science and Technology (UST), Daejeon, Republic of Korea – sequence: 12 givenname: Jae-Won surname: Huh fullname: Huh, Jae-Won organization: Department of Functional Genomics, KRIBB School of Bioscience, Korea University of Science and Technology (UST), Daejeon, Republic of Korea – sequence: 13 givenname: Sang-Rae surname: Lee fullname: Lee, Sang-Rae organization: Department of Functional Genomics, KRIBB School of Bioscience, Korea University of Science and Technology (UST), Daejeon, Republic of Korea – sequence: 14 givenname: Deog-Bon surname: Koo fullname: Koo, Deog-Bon organization: Department of Biotechnology, Daegu University, Gyeongsangbuk-do, Republic of Korea – sequence: 15 givenname: Young-Kug surname: Choo fullname: Choo, Young-Kug organization: Department of Biological Science, College of National Sciences, Wonkwang University, Jeollabuk-do, Republic of Korea – sequence: 16 givenname: Ji-Su surname: Kim fullname: Kim, Ji-Su email: kimjs@kribb.re.kr, sunuk@kribb.re.kr organization: Primate Resource Center, Korea Research Institute of Bioscience and Biotechnology, Jeollabuk-do, Republic of Korea – sequence: 17 givenname: Sun-Uk surname: Kim fullname: Kim, Sun-Uk email: kimjs@kribb.re.kr, sunuk@kribb.re.kr organization: Department of Functional Genomics, KRIBB School of Bioscience, Korea University of Science and Technology (UST), Daejeon, Republic of Korea |
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Snippet | The developmental competence of in vitro-matured oocytes is still lower than that of the in vivo-matured oocytes due to precocious meiotic resumption and... The developmental competence of in vitro -matured oocytes is still lower than that of the in vivo -matured oocytes due to precocious meiotic resumption and... |
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Title | Transient meiotic arrest maintained by DON (6-diazo-5-oxo-l-norleucine) enhances nuclear/cytoplasmic maturation of porcine oocytes |
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