The objective of this study was to determine the distribution pattern of E-cadherin in parthenogenetic sheep embryos (Ovis aries) and its relationship with fragmentation. Mature oocytes were activated with calcium ionophore (8 μg？mL？1 in TCM-199 with Hepes and 2% NCS) for 5 min, washed in TCM-199 with 20% NCS for 3 min, incubated in BO-IVC medium with 6-DMAP (2 mM) for 4 h, washed and cultured in 100 μL of the same medium at 38？C and 5% CO2. At 36 h, the development rate was assessed and stained for E-cadherin. Parthenogenetic embryos were fixed (methanol-PBS 1:1 and 2:1, 2 min each) and permeabilized in 1% Triton X100 for 5 min. Blocked in PBS with 1% albumin-fraction V for 1 h. Embryos were incubated in anti-E-cadherin primary antibody (1:50) for 24 h and in anti-IgG-FITC secondary antibody (1:50) for 24 h, determining the fluorescence intensity in the cytoplasmic region and at cell junctions in a confocal microscope. Results showed that cytoplasmic E-cadherin was highest in fragmented embryos (79.73 ± 5.3) and 4-cell embryos (57.6 ± 5.1), and lowest in 6-cell embryos (27.8 ± 3.0). At the cell junctions, progressively increased from 4-cell (45.8 ± 5.1) to 8-cell embryos (58.7 ± 6.2). It is concluded that E-cadherin exhibits a developmental stage-dependent localization with cytoplasmic predominance in early stages and a progressive accumulation at cell junctions, which coincides with that reported in embryos obtained by in vitro fertilization, while in fragmented embryos the pattern suggests alterations in the normal redistribution of E-cadherin, for example, aberrant E-cadherin localization may serve as molecular marker for poor embryo quality or impending developmental failure.
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