Effect of roscovitine and cycloheximide on ultrastructure of sheep oocytes

Crocomo, Letícia Ferrari; Filho, Wolff Camargo Marques; Sudano, Mateus José; Paschoal, D. M.; Alvarenga, Fernanda da Cruz Landim e; Bicudo, Sony Dimas

doi:10.1016/j.smallrumres.2012.07.006

Cited by 12 publications

(15 citation statements)

References 33 publications

(37 reference statements)

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Irreversible alterations such as degeneration of cumulus cells, swelling of mitochondria and reduction of cortical granules were reported by Crocomo et al. () in sheep COCs treated for 24 h with 100 μ m roscovitine. Later, maximal efficiency of meiosis inhibition was reached by Crocomo et al.…”

Section: Introductionmentioning

confidence: 62%

“…According to these same authors, the speed of development of roscovitine treated embryos was faster compared to Control. These divergent results are probably related not only to experimental conditions but also to intrinsic oocyte quality and species peculiarities (Han et al 2006;Crocomo et al 2013).…”

Section: Discussionmentioning

confidence: 99%

“…In sheep, the inhibitory potential of roscovitine as well as its effects on ultrastructure of cumulus– oocyte complexes has been recently investigated (Crocomo et al. , ,b). Irreversible alterations such as degeneration of cumulus cells, swelling of mitochondria and reduction of cortical granules were reported by Crocomo et al.…”

Section: Introductionmentioning

confidence: 99%

“…In sheep, the inhibitory potential of roscovitine as well as its effects on ultrastructure of cumulus-oocyte complexes has been recently investigated (Crocomo et al 2013(Crocomo et al , 2015a. Irreversible alterations such as degeneration of cumulus cells, swelling of mitochondria and reduction of cortical granules were reported by Crocomo et al (2013) in sheep COCs treated for 24 h with 100 lM roscovitine. Later, maximal efficiency of meiosis inhibition was reached by Crocomo et al (2015a,b) with 86.7% of oocytes kept at GV stage after treatment for 6 h with 75 lM roscovitine in serumsupplemented medium without oil overlay.…”

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

In vitro Developmental Competence of Adult Sheep Oocytes Treated with Roscovitine

Crocomo

Ariu

Bogliolo

et al. 2016

Reprod Domestic Animals

View full text Add to dashboard Cite

The efficiency of in vitro sheep embryo production is still low compared to that observed in vivo and in other species. In this context, meiotic inhibition strategies emerged as a promising alternative to improve this biotechnology. So, this study aimed to evaluate, for the first time, the effects of roscovitine on in vitro maturation of sheep oocytes and their subsequent embryo development. For this, cumulus-oocyte complexes (COCs) were cultured for 6 h in the presence (Rosco) or absence (Control) of 75 μm roscovitine and, subsequently, in vitro matured (IVM) for 18 h with gonadotropins. At 0 (Immature), 6 and 24 h of culture, the nuclear status of oocytes was evaluated by Hoechst staining. Embryo cleavage and blastocyst formation were recorded 30 h after in vitro fertilization and on day 7 of culture, respectively. Blastocyst quality was evaluated by differential staining. At 6 h, the GV rate in the Rosco treatment (93.8%) was similar to that observed in the Immature oocytes (94.9%) and significantly higher compared to Control (41.3%). After IVM for 18 h, a high and similar proportion of oocytes from Rosco (93.6%) and Control (88.4%) reached the MII stage. In both treatments, approximately 70% of oocytes cleaved and 50% of them developed up to blastocyst. The mean percentage of blastocyst cells, embryoblast, trophoblast and pyknosis did also not differ between Control and Rosco. In conclusion, roscovitine, at the studied experimental conditions, was efficient to reversibly inhibit the meiosis of adult sheep oocytes without detrimental effect on development and quality of the in vitro produced embryos.

show abstract

Section: Introductionmentioning

confidence: 62%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

In vitro Developmental Competence of Adult Sheep Oocytes Treated with Roscovitine

Crocomo

Ariu

Bogliolo

et al. 2016

Reprod Domestic Animals

View full text Add to dashboard Cite

show abstract

“…Besides, irreversible ultrastructural alterations were recorded in the cumulus cells and oocytes of sheep (Crocomo et al. ) and bovine (Lonergan et al. ) COCs treated for 24 h with 100 and 125 μ m roscovitine, respectively.…”

Section: Discussionmentioning

confidence: 99%

Effect of Oil Overlay on Inhibition Potential of Roscovitine in Sheep Cumulus‐Oocyte Complexes

Crocomo

Filho

Ulian

et al. 2015

Reprod Domestic Animals

View full text Add to dashboard Cite

Inhibitors of cyclin-dependent kinases, as roscovitine, have been used to prevent the spontaneous resumption of meiosis in vitro and to improve the oocyte developmental competence. In this study, the interference of oil overlay on the reversible arrest capacity of roscovitine in sheep oocytes as well as its effects on cumulus expansion was evaluated. For this, cumulus-oocyte complexes (COCs) were cultured for 20 h in TCM 199 with 10% foetal bovine serum (Control) containing 75 μm roscovitine (Rosco). Subsequently, they were in vitro matured (IVM) for further 18 h in inhibitor-free medium with LH and FSH. The culture was performed in Petri dishes under mineral oil (+) or in 96 well plates without oil overlay (-) at 38.5°C and 5% CO2 . At 20 and 38 h, the cumulus expansion and nuclear maturation were evaluated under stereomicroscope and by Hoechst 33342 staining, respectively. No group presented cumulus expansion at 20 h. After additional culture with gonadotrophins, a significant rate of COCs from both Control groups (+/-) exhibited total expansion while in both Rosco groups (+/-) the partial expansion prevailed. Among the oocytes treated with roscovitine, 65.2% were kept at GV in the absence of oil overlay while 40.6% of them reached MII under oil cover (p < 0.05). This meiotic arrest was reversible, and proper meiosis progression also occurred in the Control groups (+/-). So, the culture system without oil overlay improved the meiotic inhibition promoted by roscovitine without affecting the cumulus expansion rate or the subsequent meiosis progression.

show abstract