title: Microtubule distribution in somatic cell nuclear transfer bovine embryos following control of nuclear remodeling type.
abstract: This study was conducted to evaluate the microtubule distribution following control of nuclear remodeling by treatment of bovine somatic cell nuclear transfer (SCNT) embryos with caffeine or roscovitine. Bovine somatic cells were fused to enucleated oocytes treated with either 5 mM caffeine or 150 microM roscovitine to control the type of nuclear remodeling. The proportion of embryos that underwent premature chromosome condensation (PCC) was increased by caffeine treatment but was reduced by roscovitine treatment (p < 0.05). The microtubule organization was examined by immunostaining beta- and gamma-tubulins at 15 min, 3 h, and 20 h of fusion using laser scanning confocal microscopy. The gamma-tubulin foci inherited from the donor centrosome were observed in most of the SCNT embryos at 15 min of fusion (91.3%) and most of them did not disappear until 3 h after fusion, regardless of treatment (82.9-87.2%). A significantly high proportion of embryos showing an abnormal chromosome or microtubule distribution was observed in the roscovitinetreated group (40.0%, p < 0.05) compared to the caffeinetreated group (22.1%). In conclusion, PCC is a favorable condition for the normal organization of microtubules, and inhibition of PCC can cause abnormal mitotic division of bovine SCNT embryos by causing microtubule dysfunction.
authors.affiliation: College of Veterinary Medicine and Institute of Veterinary Science, Kangwon National University, Chuncheon 200-701, Korea.
authors.forename: Dae Jin
authors.initials: DJ
authors.lastname: Kwon
authors.forename: Yu Mi
authors.initials: YM
authors.lastname: Lee
authors.forename: In Sun
authors.initials: IS
authors.lastname: Hwang
authors.forename: Choon Keun
authors.initials: CK
authors.lastname: Park
authors.forename: Boo Keun
authors.initials: BK
authors.lastname: Yang
authors.forename: Hee Tae
authors.initials: HT
authors.lastname: Cheong
mesh_terms: D000818:Animals; D002110:Caffeine / Q000494:pharmacology; D002417:Cattle / Q000196:embryology / Q000502:physiology*; D002467:Cell Nucleus / Q000187:drug effects / Q000502:physiology* / Q000648:ultrastructure; D005260:Female; D005307:Fertilization in Vitro / Q000662:veterinary; D008297:Male; D018613:Microscopy, Confocal / Q000662:veterinary; D008870:Microtubules / Q000187:drug effects / Q000502:physiology* / Q000648:ultrastructure; D053652:Nuclear Transfer Techniques / Q000662:veterinary; D009865:Oocytes / Q000502:physiology*; D011247:Pregnancy; D011687:Purines / Q000494:pharmacology; D000077546:Roscovitine
chemical_list: D011687:Purines; D000077546:Roscovitine; D002110:Caffeine
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