The official journal of The Sri Lanka Veterinary Association
NUCLEAR AND MICROTUBULAR BEHAVIOR IN SOMATIC CELL SHEEP EMBRYOS ACTIVATED WITH 6-DIMETHYLAMINOPURINE AND CYCLOHEXIMIDE
Journal - Sri Lanka Veterinary Journal (volume: 62)
Volume - 62
Year - 2015
Page Number - 1
Download Article as PDF Download Full Journal as PDF
Original Article

S.L.Vet.J. 2015, 62(A): 1-6                                                                                                           

NUCLEAR AND MICROTUBULAR BEHAVIOR IN SOMATIC CELL SHEEP EMBRYOS ACTIVATED WITH 6-DIMETHYLAMINOPURINE AND CYCLOHEXIMIDE

Basil Alexander, Ph.D

Department of Farm Animal Production and Health, Faculty of Veterinary Medicine and Animal Science,
University of Peradeniya, Sri Lanka

SUMMARY: The aim of the study was to investigate the microtubules and chromatin dynamics during the first cell cycle of sheep clone embryos activated with either 6-dimethylaminopurine (6-DMAP) or cycloheximede (CHX). Fetal fibroblasts were transplanted into enucleated oocytes. The reconstructed cell-cytoplast couplets were fused and activated using ionomycin followed by culture in two separate groups containing 6-DMAP (2mM) or CHX (10 mg/ml) for 3 hours. At one hour post-fusion, 63.3% of reconstructed embryos underwent nuclear envelope break down (NEBD) and premature chromosome condensation (PCC). At one hour post-activation (hpa), a clearly visible pronucleus (diameter 6-8 μm) had already formed in the embryos treated with 6-DMAP but not in the CHX-treated group. At three hours post-activation, the 6-DMAP and CHX-treated embryos showed one swelled pronucleus with a mean diameter of 8.4 ± 1.3 μm and 25.8 ± 0.8 μm, respectively (P<0.05). In the 6-DMAP-treated embryos, most of the embryos showed a metaphase spindle as early as 16hpa but not in CHX-treated group. Typical 2 cell division was seen in most of the 6-DMAP treated embryos 28 hpa, and it was slightly delayed in the CHXtreated embryos. In summary, the artificial activating agents, 6-DMAP and CHX, exhibited different effects on chromatin remodeling, cell cycle progression rate and on the degree of pronuclear swelling which may explain the poor developmental rates and abnormal chromosome complements observed in the cloned embryos.

Corresponding author: basilalex66@gmail.com

How to cite:

Basil Alexander, (2015).Nuclear and microtubular behavior in somatic cell sheep embryos activated with 6-dimethylaminopurine and cycloheximide. Sri Lanka Veterinary Journal. 62(A), pp.1–6.

Keywords: Microtubules, Chromatin, Sheep, Embryo, Clone