Involvement of E-cadherin in early in vitro development of adult and juvenile sheep embryos

The oocyte-to-embryo transition in mammals depends on maternal proteins and transcripts, which accumulate during oocyte differentiation. The aim of the present study was to examine the role of the junctional proteins beta-catenin and E-cadherin during preimplantation in vitro embryo development in sheep, comparing the competence of adult and prepubertal oocytes. We analysed the concentration of beta-catenin and E-cadherin in immature and in vitro-matured oocytes. There was a significant increase in E-cadherin concentration after 24 h of in vitro maturation and this was lower in prepubertal oocytes than in adult ones. We therefore studied the expression and distribution of E-cadherin during the major transition from maternal to embryonic genome. E-cadherin distribution and localisation in sheep was age- and developmental-stage dependent and was related to developmental kinetics. In fact, in adults, the majority of embryos showed the proper distribution of E-cadherin just beneath the membrane surfaces of all blastomeres and the percentage of embryos with this distribution increased with the increase in cell number during development. On the contrary, and regardless of their developmental stage, the majority of prepubertal embryos showed an uneven distribution of the protein, often associated with the occurrence of cellular fragmentation. In conclusion, our results suggest that E-cadherin plays a pivotal role during preimplantation embryo growth in sheep and may be one of the possible cytoplasmic factors involved in the reduced developmental competence of prepubertal female gametes

Morphofunctional aspects of in vitro matured oocytes from prepubertal and adult sheep

The aim of this study was to analyze the morphofunctional aspects of prepubertal and adult sheep oocytes subjected to in vitro maturation (IVM). The structural and ultrastructural morphology was examined by light and transmission electron microscopy (LM and TEM) while the metabolic competence, as determined by the distribution of active mitochondria, was assessed by confocal microscopy (CM). Cumulus-oocyte complexes underwent IVM in standard condition for 24 h. Half of the mature metaphase II (MII) stage oocytes were processed for LM and TEM observations. The other half was subjected to immunostaining with MitoTracker Red (to stain mitochondria with functionally active membrane potential), SNARF-1 (a pH sensitive fluoroprobe), Hoechst 33342 (to label DNA) and analyzed by CM. Immature germinal vesicle (GV) stage oocytes, retrieved at 0 h IVM, were used as controls. By LM and TEM all the oocytes were regularly rounded, covered by microvilli and surrounded by an intact zona pellucida. Numerous rounded, oval or hooded mitochondria appeared either isolated or grouped in the ooplasm. The GV was usually rounded in prepubertal oocytes. In the adult, the GV often appeared flattened against the oolemma, with a crescent-shaped outline (an early sign of meiotic resumption). Scattered cortical granules (CGs) were rarely found in the ooplasm of both prepubertal and adult GV oocytes. After 24 h of IVM, CGs became abundant and distributed in a single row under the oolemma, particularly in adult oocytes. CM showed a homogeneous fine-to-granular mitochondrial distribution in prepubertal GV and MII oocytes. In adult GV oocytes, the mitochondrial distribution pattern was granular while in MII oocytes mitochondria were arranged in heterogeneous clusters. Thus, both prepubertal and adult oocytes completed maturation after 24 h in culture and showed an overall good preservation after IVM. However, the diverse distribution patterns of mitochondria in prepubertal oocytes reflect their low developmental competence

Warm Nuclei: Nuclear Structure Effects on the Order-to-Chaos Transition Region

International audienceThe \gamma -decay from the warm rotation in the transition region between order and chaos is studied in the superdeformed (SD) nuclei 151Tb and 196Pb, using the EUROBALL IV array. A number of observables, testing the decay dynamics in the SD well, are compared with predictions from a Monte Carlo simulation of the \gamma -decay based on microscopic calculations of discrete levels and decay probabilities. Agreement with the data is found only assuming an enhancement of the B(E1) strength around 1 MeV by a factor of 10-100, which is consistent with the evidence for octupole vibrations in both mass regions. The work shows the relevance of \gamma -spectroscopy in the order-to-chaos regime to highlight specific nuclear structure effects