G6PDH-activity in equine oocytes correlates with morphology, expression of candidate genes for viability, and preimplantative in vitro development.
Abstract: Efficiencies for in vitro production of equine embryos are still low due to highly variable developmental competences of equine immature oocytes. In contrast to the equine, in vitro developmental competence of immature oocytes has been predicted successfully by the activity of glucose-6-phosphate dehydrogenase (G6PDH) indicated by brilliant cresyl blue (BCB) dye in a range of different species. Therefore, the aim of the present study was to test the association between G6PDH activity in equine oocytes with: (1) cumulus morphology and oocyte properties in terms of diameter and volume; (2) maturational competence; (3) gene expression of certain molecular markers; and (4) in vitro embryo development after intracytoplasmic sperm injection. Equine oocytes were exposed to BCB stain and were classified as BCB+ or BCB- according to their ability to convert the dye from blue to colorless. Additionally, BCB+ and BCB- oocytes were subclassified as having a compact (Cp) or expanded (Ex) cumulus complex. As a result, BCB+ oocytes had a greater proportion of expanded cumulus oocyte complexes compared with BCB- oocytes (71.2% vs. 49.5%). Moreover, we observed a significant difference in oocyte diameter and volume between BCB+ and BCB- oocytes irrespective of cumulus morphology. BCB+ oocytes reached a higher maturation rate compared with BCB- oocytes (59.0% vs. 28.7%). Regarding the analyzed candidate genes, relative transcript abundance was significantly different for nine genes. The expression of eight genes was significantly higher (P < 0.05) for BCB+ oocytes, including ATPV6E, IF-3, TFAM, DNMT1, STAT3, Aurora-A, ODC1, and CKS2 whereas BCB- oocytes showed higher in expression of COX1. These results are in line with the observed developmental competence. Cleavage rate (45.9% vs. 29.0%) and percentage of embryos that reached the blastocyst stage (9.2% vs. 1.4%) were significantly higher for embryos derived from BCB+ oocytes compared with BCB- oocytes. In conclusion, the present study provides evidence that G6PDH-activity in immature equine oocytes is a useful predictor for subsequent in vitro developmental competence.
Copyright © 2011 Elsevier Inc. All rights reserved.
Publication Date: 2011-08-05 PubMed ID: 21820165DOI: 10.1016/j.theriogenology.2011.05.025Google Scholar: Lookup
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Summary
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The research explores the connection between glucose-6-phosphate dehydrogenase (G6PDH) activity in equine oocytes and their in vitro development competence. The study finds that G6PDH-activity can be a useful predictor of an oocyte’s subsequent in vitro developmental capability.
Research Purpose and Methodology
- The overall aim of the research was to analyze the relationship between G6PDH activity in equine oocytes (immature horse eggs) and four key aspects: cumulus morphology, oocyte diameter and volume, maturation competence, gene expression of certain molecular markers, and in vitro embryo development post intracytoplasmic sperm injection.
- In the course of the study, equine oocytes were exposed to a blue dye known as Brilliant Cresyl Blue (BCB). The oocytes were then categorized into two groups: those that changed the dye from blue to colorless (BCB+) and those that did not (BCB-).
- Further, they divided these into subcategories based on their cumulus complexes – those having compact (Cp) or expanded (Ex) forms.
Detailed Findings
- The BCB+ oocytes demonstrated a higher proportion of expanded cumulus oocyte complexes compared to BCB- oocytes (71.2% vs. 49.5%).
- It was observed that there was a significant difference in the size and volume of the oocytes between the BCB+ and BCB- groups, regardless of cumulus morphology.
- Unsurprisingly, BCB+ oocytes exhibited a higher rate of maturation compared to BCB- oocytes (59.0% vs. 28.7%).
- In terms of gene expression, the study found significant differences for nine genes. BCB+ oocytes showed a higher level of expression for eight genes (ATPV6E, IF-3, TFAM, DNMT1, STAT3, Aurora-A, ODC1, and CKS2) while one gene (COX1) was expressed more in BCB- oocytes.
- Furthermore, cleavage rate and the percentage of embryos reaching the blastocyst stage were significantly greater in BCB+ oocytes (45.9% vs. 29.0% for cleavage rate, and 9.2% vs. 1.4% reaching the blastocyst stage).
Conclusion
- The research concludes that the activity of G6PDH in equine oocytes is a valuable predictor for subsequent in vitro developmental competence.
- This suggests that monitoring G6PDH-activity could potentially improve the efficiency of in vitro production of equine embryos, which currently suffers from considerable variability in developmental competences between individual oocytes.
Cite This Article
APA
Mohammadi-Sangcheshmeh A, Held E, Ghanem N, Rings F, Salilew-Wondim D, Tesfaye D, Sieme H, Schellander K, Hoelker M.
(2011).
G6PDH-activity in equine oocytes correlates with morphology, expression of candidate genes for viability, and preimplantative in vitro development.
Theriogenology, 76(7), 1215-1226.
https://doi.org/10.1016/j.theriogenology.2011.05.025 Publication
Researcher Affiliations
- Institute of Animal Science, Animal Breeding and Husbandry Group, University of Bonn, Bonn, Germany.
MeSH Terms
- Animals
- Cumulus Cells / cytology
- Embryonic Development
- Female
- Fertilization in Vitro / veterinary
- Glucosephosphate Dehydrogenase / metabolism
- Horses
- Oocytes / cytology
- Oocytes / metabolism
Citations
This article has been cited 10 times.- Pollard CL, Younan A, Swegen A, Gibb Z, Grupen CG. Insights into the NAD(+) biosynthesis pathways involved during meiotic maturation and spindle formation in porcine oocytes. J Reprod Dev 2022 Jun 1;68(3):216-224.
- Srirattana K, Kaneda M, Parnpai R. Strategies to Improve the Efficiency of Somatic Cell Nuclear Transfer. Int J Mol Sci 2022 Feb 10;23(4).
- Piras AR, Ariu F, Zedda MT, Paramio MT, Bogliolo L. Selection of Immature Cat Oocytes with Brilliant Cresyl Blue Stain Improves In Vitro Embryo Production during Non-Breeding Season. Animals (Basel) 2020 Aug 24;10(9).
- Heydari S, Eidi A, Kouhkan F, Tvrda E, Mohammadi-Sangcheshmeh A. Effects of increasing lipopolysaccharide concentrations on in vitro developmental competence of ovine oocytes. Anim Reprod 2020 Jul 7;17(2):e20190125.
- Piras AR, Menéndez-Blanco I, Soto-Heras S, Catalá MG, Izquierdo D, Bogliolo L, Paramio MT. Resveratrol supplementation during in vitro maturation improves embryo development of prepubertal goat oocytes selected by brilliant cresyl blue staining. J Reprod Dev 2019 Apr 12;65(2):113-120.
- Karami Shabankareh H, Azimi G, Torki M. Developmental competence of bovine oocytes selected based on follicle size and using the brilliant cresyl blue (BCB) test. Iran J Reprod Med 2014 Nov;12(11):771-8.
- Ashry M, Lee K, Mondal M, Datta TK, Folger JK, Rajput SK, Zhang K, Hemeida NA, Smith GW. Expression of TGFβ superfamily components and other markers of oocyte quality in oocytes selected by brilliant cresyl blue staining: relevance to early embryonic development. Mol Reprod Dev 2015 Mar;82(3):251-64.
- Mohammadi-Sangcheshmeh A, Veshkini A, Hajarizadeh A, Jamshidi-Adegani F, Zhandi M, Abazari-Kia AH, Cinar MU, Soleimani M, Gastal EL. Association of glucose-6-phosphate dehydrogenase activity with oocyte cytoplasmic lipid content, developmental competence, and expression of candidate genes in a sheep model. J Assist Reprod Genet 2014 Aug;31(8):1089-98.
- Abazari-Kia AH, Mohammadi-Sangcheshmeh A, Dehghani-Mohammadabadi M, Jamshidi-Adegani F, Veshkini A, Zhandi M, Cinar MU, Salehi M. Intracellular glutathione content, developmental competence and expression of apoptosis-related genes associated with G6PDH-activity in goat oocyte. J Assist Reprod Genet 2014 Mar;31(3):313-21.
- Pawlak P, Warzych E, Chabowska A, Lechniak D. Differences in cytoplasmic maturation between the BCB+ and control porcine oocytes do not justify application of the BCB test for a standard IVM protocol. J Reprod Dev 2014 Mar 7;60(1):28-36.
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