Vitrification of bovine oocytes at different meiotic stages using the Cryotop method: Assessment of morphological, molecular and functional patterns

Sprícigo, J. F. W.; Morais, K.; Ferreira, A. R.; Machado, G. M.; Gomes, Ana C.; Rumpf, R.; Franco, M. M.; Dode, M. A. N.

doi:10.1016/j.cryobiol.2014.07.015

Cited by 29 publications

(23 citation statements)

References 59 publications

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“…BRK1 gene is mainly involved in regulation of actin and microtubule organization. Together, these results explain the disruption of the cytoskeleton (Dobrinsky, Pursel, Long, & Johnson, ; Hwang & Hochi, ; Vincent, Pickering, Johnson, & Quick, ) and cytoplasm (Sprícigo et al., ) in vitrified mammalian oocytes.…”

Section: Discussionmentioning

confidence: 82%

Effect of vitrification on the mRNA transcriptome of bovine oocytes

Wang

et al. 2017

Reprod Domestic Animals

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Vitrification has been shown to decrease the developmental capacity of mammalian oocytes, and this is closely associated with the abnormal mRNA expressions of vitrified oocytes. However, the effect of vitrification on transcriptional machinery of oocytes examined by RNA sequencing (RNA-seq) has yet to be defined. In the present study, the mRNA transcriptomes of fresh and vitrified bovine oocytes were analysed by Smart-seq2 with the differently expressed genes determined by DEseq2 (an adjusted p-value of .05 and a minimum fold change of 2). The differentially expressed mRNAs were then searched against the Gene Ontology (GO) and Genomes (KEGG) database. Finally, the mRNA expressions of 10 candidate genes were validated using quantitative real-time PCR (qRT-PCR). Approximately 12,000 genes were detected in each sample of fresh or vitrified oocytes. Of these, the expression levels of 102 genes differed significantly in vitrified groups: 12 genes mainly involved in cell cycle, fertilization and glucose metabolism were upregulated, and 90 genes mainly involved in mitochondria, ribosomal protein, cytoskeleton, transmembrane protein, cell cycle and calcium ions were downregulated. GO analysis showed that these genes were mainly enriched in terms of membrane-bounded organelles, macromolecular complex, and intracellular part. The mRNA expression levels of 10 candidate genes selected randomly were in agreement with the results of the RNA-seq. In conclusion, our results showed that vitrification affected the mRNA transcriptome of bovine oocytes by downregulating genes, which contributed to the decreased developmental capacity of vitrified oocytes. Our findings will be useful in determining approaches to improve the efficiency of vitrified oocytes.

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Section: Discussionmentioning

confidence: 82%

Effect of vitrification on the mRNA transcriptome of bovine oocytes

Wang

et al. 2017

Reprod Domestic Animals

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“…These differences in results may be due to differences in samples of studying. Other studies have also reported damages to the fibers of the cytoskeleton and microtubules resulting from vitrification (Shi et al, 2007;Nottola et al, 2014;Sprícigo et al, 2014).…”

Section: Discussionmentioning

confidence: 99%

Assessment of Mouse Oocytes Ultrastructure Following Vitrification Before and After in vitro Maturation

et al. 2018

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IZADI, M.; EFTEKHAR, V. S. H.; AKBARI, H.; ASADI, S. M. & MOKHTARI, T.Assessment of mouse oocytes ultrastructure following vitrification before and after in vitro maturation. Int. J. Morphol., 36(1):180-188, 2018. SUMMARY:Vitrification is a physical process in which the concentrated cryoprotectant solution after exposure to extreme cold without ice crystal formation in living cells to be converted glassing state. In this study, maturation rate and ultrastructure of mouse oocytes followed by vitrification before or after in-virto maturation (IVM) were evaluated. A total of 373 germinal vesicle oocytes were obtained from ovaries and divided into three fresh IVM, IVM vitrified, vitrified IVM groups. Ten metaphase II oocytes were obtained from uterine tubes and considered as the control group. Oocytes in vitrified groups were vitrified by Cryotop using vitrification medium and kept in liquid nitrogen. The maturation media was a-MEM supplemented with rFSH + hCG. After 24-48 h of incubation, the oocytes were investigated for nuclear maturation and ultrastructural changes using transmission electron microscopy (TEM). The oocyte maturation rate in vIVM group was significantly lower than IVMv group, when the two groups were compared with vIVM had the highest maturity. The evaluation ultrastructure of the four groups showed that the number of cortical granules, microvilli and mitochondria-SER aggregates in vIVM group were lowest and the highest amongst the number of vacuoles. Zona pellucida was darker than the control group in two freeze groups vIVM and IVMv. Most similar groups to the control group were group vIVM, Group IVMv and ultimately vIVM group, respectively. According to the results, IVM procedure is more efficient when it is performed before oocyte vitrification.

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“…For example, in bovine embryos produced in vivo, less amino acid is produced than in IVP embryos (STURMEY et al, 2010 (GUERIF et al, 2013).…”

Section: Embryonic Energy Metabolismmentioning

confidence: 99%

“…Additionally, many studies have been carried out to identify genes of interest, according to the literature of the field (OROZCO-LUCERO et al, 2014;SPRICIGO et al, 2014;LEME et al, 2016). In the case of IVP bovine embryos, it is known that the expression of genes important for embryonic development is different from those produced in vivo (MUNDIM et al, 2009).…”

Section: Genetic Markers Related To Lipid Metabolismmentioning

confidence: 99%

Change in energy metabolism of in vitro produced embryos: an alternative to make them more cryoresistant?

Dias

Pivato

Dode

2017

SCA

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For the development of in vitro produced (IVP) as well as in vivo produced bovine embryos, it is extremely important that their energy metabolism works properly because the embryo must be able to metabolize energy substrates that are necessary for producing energy. Lipids play an important role in early embryonic development, acting as source of energy for oocytes and embryos. However, it is known that oocytes and embryos, mainly IVP, accumulate large amounts of lipids in the cytoplasm. Although they are extremely important in embryonic development, lipids have been associated with the reduced survival of bovine embryos following cryopreservation. There is evidence that at least four different categories of lipids affect embryo survival after cryopreservation, including triglycerides (TAG), free fatty acids, cholesterol and phospholipids. Thus, many studies are being conducted to improve the resistance of IVP embryos to the cryopreservation process by reducing the concentration or removing the source of serum from the medium or by reducing oocyte/embryo lipids using mechanical or chemical means. Regarding the use of delipidating agents that reduce the uptake and synthesis of fatty acids (FA) by cells, substances such as phenazine ethosulfate (PES), forskolin, L-carnitine and isomers of conjugated linoleic acid (CLA) have been utilized. This review aims to address important issues related to embryonic energy metabolism, the importance of lipid metabolism and its relation to the cryopreservation of IVP bovine embryos by summarizing the latest research in this field. Key words: Bovine embryos. Cryopreservation. Energy metabolism. Lipids. ResumoPara o desenvolvimento de embriões bovinos produzidos in vitro (PIV) e in vivo, é extremamente importante que o metabolismo energético funcione de maneira adequada, pois o embrião precisa ser capaz de metabolizar os substratos energéticos necessários para a produção de energia. Os lipídios desempenham papel importante no desenvolvimento embrionário inicial, atuando como fonte de energia para oócitos e embriões. Entretanto, sabe-se que oócitos e embriões bovinos, principalmente os PIV, acumulam grande quantidade de lipídios no citoplasma. Apesar de serem extremamente importantes no desenvolvimento embrionário, os lipídios têm sido associados com a reduzida sobrevivência após a criopreservação de embriões bovinos. Existem evidências de que pelo menos quatro classes de lipídios afetam a sobrevivência embrionária pós-criopreservação, sendo os triglicerídeos (TAG), ácidos graxos livres, colesterol e os fosfolipídios. Sendo assim, vários estudos estão sendo realizados com o intuito de melhorar a resistência dos embriões PIV ao processo de criopreservação, realizando a redução ou retirada do soro do meio ou a redução mecânica ou química de lipídios. Com relação ao uso de agentes delipidantes que diminuam a captação e síntese de ácidos graxos (AG) pelas células, substâncias como fenazina etossulfato (PES), forskolin, L-carnitina e isômeros do ácido linoleico conjugado (CLA) t...

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Vitrification of bovine oocytes at different meiotic stages using the Cryotop method: Assessment of morphological, molecular and functional patterns

Cited by 29 publications

References 59 publications

Effect of vitrification on the mRNA transcriptome of bovine oocytes

Effect of vitrification on the mRNA transcriptome of bovine oocytes

Assessment of Mouse Oocytes Ultrastructure Following Vitrification Before and After in vitro Maturation

Change in energy metabolism of in vitro produced embryos: an alternative to make them more cryoresistant?

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