Early development of cloned bovine embryos produced from oocytes enucleated by fluorescence metaphase II imaging using a conventional halogen-lamp microscope
| Autor: | Iwamoto, D., Yamagata, K., Kishi, M., Hayashi-Takanaka, Y., Kimura, Hiroshi, Wakayama, T., Saeki, K. |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2015 |
| Předmět: |
Male
Nuclear Transfer Techniques Immunoconjugates Cloning Organism Enucleation Embryonic Development Fertilization in Vitro Biology Chromosomes Fluorescence Phycoerythrin/chemistry Oocytes/*cytology Nuclear Transfer Techniques/*veterinary medicine Animals Blastocyst Metaphase Original Research Cloning Chromosome Embryo Phycoerythrin Cloning Organism/*methods Cell Biology Oocyte Molecular biology Cell biology medicine.anatomical_structure Immunoconjugates/chemistry Oocytes Somatic cell nuclear transfer Cattle Blastocyst/*cytology Developmental Biology Biotechnology Fertilization in Vitro/veterinary |
| Zdroj: | Cellular reprogramming. 17(No. 2):106-14 |
| ISSN: | 2152-4971 |
| Popis: | Enucleation of a recipient oocyte is one of the key processes in the procedure of somatic cell nuclear transfer (SCNT). However, especially in bovine species, lipid droplets spreading in the ooplasm hamper identification and enucleation of metaphase II (MII) chromosomes, and thereby the success rate of the cloning remains low. In this study we used a new experimental system that enables fluorescent observation of chromosomes in living oocytes without any damage. We succeeded in visualizing and removing the MII chromosome in matured bovine oocytes. This experimental system consists of injecting fluorescence-labeled antibody conjugates that bind to chromosomes and fluorescent observation using a conventional halogen-lamp microscope. The cleavage rates and blastocyst rates of bovine embryos following in vitro fertilization (IVF) decreased as the concentration of the antibody increased (p0.05). For the production of SCNT embryos, the new system can be used as a reliable predictor of the location of metaphase plates in opaque oocytes, such as those in ruminant animals. |
| Databáze: | OpenAIRE |
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