title: Effect of demecolcine and nocodazole on the efficiency of chemically assisted removal of chromosomes and the developmental potential of nuclear transferred porcine oocytes.
abstract: Brief treatment of metaphase II (MII) stage porcine oocytes with 0.4 microg/mL demecolcine in the presence of 0.05 M sucrose produces a membrane protrusion that contains a condensed chromosome mass. The present study examined the optimal conditions for demecolcine and nocodazole treatment in chemically assisted removal of chromosomes. When matured oocytes were treated with 0.1-0.4 microg/mL demecolcine for 60 min or with 0.4 microg/mL demecolcine for 30 min or 3 microg/mL nocodazole for 30 or 60 min, more than 70% of oocytes had a membrane protrusion containing condensed chromosomes were located. There was no difference in the in vitro developmental potential of enucleated oocytes assisted by 0.1 and 0.4 microg/mL demecolcine or 3 microg/mL nocodazole that received porcine somatic cells. After transfer to 10 recipients, however, two of six recipients that received demecolcine-treated enucleated eggs produced four healthy cloned piglets, but none of the four recipients of nocodazole-treated enucleated eggs produced piglets. Further studies are required to increase the successful development to term because the proportion of live piglets was low (4/2, 672, 0.15%).
authors.affiliation: Laboratory of Animal Reproduction, College of Agriculture, Kinki University, 3327-204, Nakamachi, Nara 631-8505, Japan.
authors.forename: Masahiro
authors.initials: M
authors.lastname: Kawakami
authors.forename: Tetsuya
authors.initials: T
authors.lastname: Tani
authors.forename: Akiko
authors.initials: A
authors.lastname: Yabuuchi
authors.forename: Tatsuya
authors.initials: T
authors.lastname: Kobayashi
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Murakami
authors.forename: Tatsuya
authors.initials: T
authors.lastname: Fujimura
authors.forename: Yoko
authors.initials: Y
authors.lastname: Kato
authors.forename: Yukio
authors.initials: Y
authors.lastname: Tsunoda
mesh_terms: D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology*; D002467:Cell Nucleus / Q000235:genetics; D002478:Cells, Cultured; D002875:Chromosomes / Q000187:drug effects* / Q000235:genetics; D019976:Cloning, Organism / Q000379:methods*; D003703:Demecolcine / Q000494:pharmacology*; D004624:Embryo Transfer; D005314:Embryonic and Fetal Development / Q000187:drug effects*; D005260:Female; D005307:Fertilization in Vitro; D008297:Male; D015739:Nocodazole / Q000494:pharmacology*; D009865:Oocytes / Q000187:drug effects* / Q000502:physiology; D011247:Pregnancy; D013552:Swine
chemical_list: D000970:Antineoplastic Agents; D015739:Nocodazole; D003703:Demecolcine