The utility of Brilliant Cresyl Blue (BCB) staining of mammalian oocytes used for in vitro embryo production (IVP)

Opiela, Jolanta; Kątska-Książkiewicz, L.

doi:10.1016/j.repbio.2013.07.004

Cited by 41 publications

(37 citation statements)

References 35 publications

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“…However, the predictive value of the BCB approach used in these studies is still controversial [36]. Therefore, it is still too early to apply the BCB tool in human ARTs and further experimental studies in animal model are needed to clarify whether the method is generally applicable.…”

Section: Discussionmentioning

confidence: 99%

Intracellular glutathione content, developmental competence and expression of apoptosis-related genes associated with G6PDH-activity in goat oocyte

Abazari-Kia

Mohammadi‐Sangcheshmeh

Dehghani-Mohammadabadi

et al. 2013

J Assist Reprod Genet

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Purpose To associate glucose-6-phosphate dehydrogenase (G6PDH) activity in goat oocytes with intracellular glutathione (GSH) content, meiotic competence, developmental potential, and relative abundance of Bax and Bcl-2 genes transcripts. Methods Goat oocytes were exposed to brilliant cresyl blue (BCB) staining test and categorized into BCB + (bluecytoplasm), and BCB − (colorless-cytoplasm) groups. A group of oocytes were not exposed to BCB test and was considered as a control group. After maturation in vitro, a group of oocytes were used for determination of nuclear status and intracellular GSH content while another group was subjected to parthenogenetic activation followed by in vitro embryo culture.Results We found that BCB + oocytes not only yielded higher rate of maturation, but also showed an increased level of intracellular GSH content than BCB − and control oocytes.Furthermore, BCB + oocytes produced more blastocysts than BCB − and control oocytes. Our data revealed that the expression of anti-apoptotic (Bcl-2) and pro-apoptotic (Bax) genes were interacted with G6PDH-activity in mature oocyte, their surrounding cumulus cells, and blastocyst-stage embryos.Conclusions The results of this study demonstrate that selection of goat oocytes based on G6PDH-activity through the BCB test improves their developmental competence, increases intracellular GSH content, and affects the expression of the apoptosis-related genes.

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Section: Discussionmentioning

confidence: 99%

Intracellular glutathione content, developmental competence and expression of apoptosis-related genes associated with G6PDH-activity in goat oocyte

Abazari-Kia

Mohammadi‐Sangcheshmeh

Dehghani-Mohammadabadi

et al. 2013

J Assist Reprod Genet

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“…Pawlak et al [21] observed a higher rate of chromosomal aberrations in porcine oocytes after BCB staining. A higher susceptibility to apoptosis of BCB-stained bovine oocytes as revealed by the relative transcript abundance of BCL-2 and BAX genes was noticed by Opiela et al [9, 22]. Wongsrikeao et al [2] reported that BCB staining after IVM had a negative impact on the cleavage and development of porcine embryos.…”

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confidence: 92%

Differences in Cytoplasmic Maturation Between the BCB+ and Control Porcine Oocytes Do Not Justify Application of the BCB Test for a Standard IVM Protocol

et al. 2014

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The Brilliant Cresyl Blue (BCB) test relies on G6PDH activity and a simple protocol for the selection of higher quality oocytes. Although the BCB+ oocytes of all the species that have been investigated are characterized by superior quality when compared to BCB- counterparts, application of the test for embryo production still remains an open issue. The aim of our study was to compare BCB+ and the control oocytes (not subjected to the BCB test) in terms of selected aspects of cytoplasmic maturation (mtDNA copy number, mitochondria distribution, relative transcript abundance of six marker genes). The results of our study revealed more relevant differences within the BCB+ and the control oocytes (before and after IVM) than between the two categories of oocytes. There was no difference in the transcript abundance of the BCB+ and the control oocytes in 5 out of 6 analyzed genes (BMP15, GDF9, ATP5A1, EEF1A, ZAR1) and in mtDNA content (pre-IVM 179609 vs. 176595 and post-IVM 187243 vs. 246984, respectively). With regard to mitochondria distribution in pre- and post-IVM oocytes, there was nonsignificant tendency for a more frequent occurrence of the expected patterns in the BCB+ group. The results of the present study do not support the application of BCB staining in a routine IVM protocol due to relatively high similarity in selected parameters characterizing cytoplasmic maturation of BCB+ and control oocytes. This high similarity may results from the limited amount of less competent BCB- oocytes (10%) still present among nonselected oocytes of proper morphology.

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“…Despite recent advances in reproductive and developmental biology, morphological criteria remain the most widely used method for oocyte selection because molecular marker (RNA)-based evaluation of oocytes is invasive, costly, and/or not readily amenable to rapid classification. Brilliant cresyl blue (BCB) staining is a non-invasive method used to select competent oocytes in different species, including cattle, goats, sheep, pigs (Opiela and Katska-Ksiazkiewicz, 2013), buffalo (Manjunatha et al, 2007), horses (Mohammadi-Sangcheshmeh et al, 2011;Pereira et al, 2014), and mice (Wu et al, 2007). BCB is used to determine the intracellular activity of glucose-6-phosphate dehydrogenase (G6PDH, Alm et al, 2005), a regulatory enzyme that is synthesized and accumulates during oocyte growth and whose activity gradually decreases as oocytes complete their growth phase (Mangia and Epstein, 1975).…”

Section: Introductionmentioning

confidence: 99%

Expression of TGFβ superfamily components and other markers of oocyte quality in oocytes selected by brilliant cresyl blue staining: Relevance to early embryonic development

Ashry

Lee

Mondal

et al. 2015

Molecular Reproduction Devel

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Brilliant cresyl blue (BCB) is a super vital stain that has been used to select competent oocytes in different species. The objectives of present studies were to determine mRNA abundance for select TGFβ superfamily components, SMAD2/3 and SMAD1/5 phosphorylation levels and transcript abundance for other oocyte (JY1) and cumulus cell (CTSB, CTSK, CTSS and CTSZ) markers of oocyte quality in bovine oocytes and or adjacent cumulus cells classified based on developmental potential using BCB staining. The ability of exogenous FST, JY1, or cathepsin inhibitor treatment to enhance development of embryos derived from poor quality oocytes selected based on BCB staining was also determined. Cumulus oocyte complexes (COCs) from abattoir derived ovaries were subjected to BCB staining and GV stage oocytes and cumulus cells harvested from control, BCB+ and BCB- (poor oocyte quality) groups for real time PCR or Western blot analysis. Remaining COCs underwent in vitro maturation, in vitro fertilization and embryo culture in presence or absence of above described treatments. Levels of FST, JY1, BMP15 and SMAD1, 2, 3 and 5 transcripts were higher in BCB+ oocytes whereas abundance of CTSB, CTSK, CTSS and CTSZ mRNAs was higher in cumulus cells surrounding poor quality BCB- oocytes. Western blot analysis revealed SMAD1/5 and SMAD2/3 phosphorylation were higher in BCB+ than BCB− oocytes. Embryo culture studies demonstrated that follistatin and cathepsin inhibitor treatment but not JY-1 treatment can promote developmental competence of BCB- oocytes. Results provide further understanding of molecular indices of oocyte competence.

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The utility of Brilliant Cresyl Blue (BCB) staining of mammalian oocytes used for in vitro embryo production (IVP)

Cited by 41 publications

References 35 publications

Intracellular glutathione content, developmental competence and expression of apoptosis-related genes associated with G6PDH-activity in goat oocyte

Intracellular glutathione content, developmental competence and expression of apoptosis-related genes associated with G6PDH-activity in goat oocyte

Differences in Cytoplasmic Maturation Between the BCB+ and Control Porcine Oocytes Do Not Justify Application of the BCB Test for a Standard IVM Protocol

Expression of TGFβ superfamily components and other markers of oocyte quality in oocytes selected by brilliant cresyl blue staining: Relevance to early embryonic development

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