Protective effects of the cumulus-corona radiata complex during vitrification of horse oocytes.
Abstract: Vitrifying oocytes is a potentially valuable means of preserving the female germ line, but significantly compromises oocyte developmental competence. This study examined the hypothesis that the cumulus complex protects the oocyte during vitrification. Vitrified-warmed immature cumulus oocyte complexes (COCs) were labelled with a plasma membrane impermeant DNA marker (ethidium homodimer-1) to examine the percentage and location of dead cumulus cells, and to investigate the effect of the proportion of dead cells (+1,+2 or +3) on the success of in vitro maturation (IVM). Further, oocytes were labelled for connexin-43 or injected with Lucifer yellow dye to determine whether the integrity of the gap junctions between an oocyte and its cumulus was compromised by vitrification. Finally, the effect of denuding immature and mature oocytes on their ability to withstand vitrification was examined. Cryopreserving immature COCs increased the number of dead cumulus cells (13 vs 2.6% for controls; P<0.05). However, an increased proportion of dead cumulus cells did not affect post-warming maturation rates (approximately 30% MII) presumably because dead cells were located at the periphery of the cumulus mass and cumulus-oocyte gap junction communication was not disrupted. Moreover, cumulus removal prior to IVM or vitrification indicated that while the cumulus does protect immature oocytes during vitrification it does so by mechanisms other than support during maturation. Cumulus presence was also found to protect mature equine oocytes against vitrification-induced damage since cumulus-enclosed MII oocytes preserved their meiotic spindle quality better during vitrification than denuded oocytes (38.1 vs 3.1% normal spindles; P<0.05).
Publication Date: 2008-12-10 PubMed ID: 19073713DOI: 10.1530/REP-08-0333Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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This research investigates how the cumulus oocyte complex (COC) protects horse oocytes against damage during vitrification, a preservation method with potential for conserving the female germ line. It was found that the cumulus helps preserve the developmental competence of the oocyte not only during maturation but also throughout the vitrification process.
Objective of the Research
- The purpose of this study was to examine the potential protective effects of the cumulus-corona radiata complex, particularly during the vitrification of horse oocytes (immature egg cells). Vitrification is a preservation technique that can be invaluable for safeguarding the female germ line, despite its notable impact on the developmental competence of oocytes.
Conducting the Research
- The researchers vitrified-warmed immature COCs and labelled the cells with a DNA marker to examine the percentage and location of dead cumulus cells. They also examined if the proportion of dead cells affected the success of in vitro maturation (IVM).
- They used a dye and labelling procedure to determine if vitrification disrupted the integrity of gap junctions between an oocyte and its cumulus.
- The team tested the effect of denuding (removing layers of cumulus cells) immature and mature oocytes on their ability to endure vitrification.
Findings of the Study
- Cryopreserving immature COCs through vitrification increased the number of dead cumulus cells. However, this did not affect post-warming maturation rates most likely because the dead cells were on the periphery and didn’t interfere with the gap junction communication.
- The research revealed that though the cumulus does indeed protect immature oocytes during vitrification, it does so through mechanisms not directly related to maturation support.
- The presence of the cumulus correlated with improved resilience of mature equine oocytes to vitrification-induced damage. Cumulus-enclosed mature oocytes better preserved their meiotic spindle quality during vitrification than did denuded oocytes.
Implications of the Research
- This study provides valuable insights into how the cumulus complex protects oocytes during vitrification. Understanding this can direct the development of better preservation protocols, ultimately promoting the conservation of the female germ line.
Cite This Article
APA
Tharasanit T, Colleoni S, Galli C, Colenbrander B, Stout TA.
(2008).
Protective effects of the cumulus-corona radiata complex during vitrification of horse oocytes.
Reproduction, 137(3), 391-401.
https://doi.org/10.1530/REP-08-0333 Publication
Researcher Affiliations
- Department of Equine Sciences, Faculty of Veterinary Medicine, Utrecht University, Utrecht, The Netherlands.
MeSH Terms
- Animals
- Cell Survival
- Cells, Cultured
- Cryopreservation / methods
- Cryopreservation / veterinary
- Female
- Gap Junctions / physiology
- Horses
- Meiosis
- Microscopy, Confocal
- Oocytes / cytology
Citations
This article has been cited 12 times.- Temerario L, Monaco D, Mastrorocco A, Martino NA, Cseh S, Lacalandra GM, Ciani E, Dell'Aquila ME. New Strategies for Conservation of Gentile di Puglia Sheep Breed, an Autochthonous Capital of Millennial Tradition in Southern Italy. Animals (Basel) 2023 Jul 20;13(14).
- Angel-Velez D, Meese T, Hedia M, Fernandez-Montoro A, De Coster T, Pascottini OB, Van Nieuwerburgh F, Govaere J, Van Soom A, Pavani K, Smits K. Transcriptomics Reveal Molecular Differences in Equine Oocytes Vitrified before and after In Vitro Maturation. Int J Mol Sci 2023 Apr 7;24(8).
- Angel-Velez D, De Coster T, Azari-Dolatabad N, Fernandez-Montoro A, Benedetti C, Bogado Pascottini O, Woelders H, Van Soom A, Smits K. New Alternative Mixtures of Cryoprotectants for Equine Immature Oocyte Vitrification. Animals (Basel) 2021 Oct 28;11(11).
- López A, Betancourt M, Ducolomb Y, Rodríguez JJ, Casas E, Bonilla E, Bahena I, Retana-Márquez S, Juárez-Rojas L, Casillas F. DNA damage in cumulus cells generated after the vitrification of in vitro matured porcine oocytes and its impact on fertilization and embryo development. Porcine Health Manag 2021 Oct 18;7(1):56.
- Sowińska N, Zahmel J, Niżański W, Hribal R, Fernandez-Gonzalez L, Jewgenow K. Meiotic Status Does Not Affect the Vitrification Effectiveness of Domestic Cat Oocytes. Animals (Basel) 2020 Aug 7;10(8).
- Ishii T, Tomita K, Sakakibara H, Ohkura S. Embryogenesis of vitrified mature bovine oocytes is improved in the presence of multi-layered cumulus cells. J Reprod Dev 2018 Feb 27;64(1):95-99.
- Khalili MA, Shahedi A, Ashourzadeh S, Nottola SA, Macchiarelli G, Palmerini MG. Vitrification of human immature oocytes before and after in vitro maturation: a review. J Assist Reprod Genet 2017 Nov;34(11):1413-1426.
- Purohit GN, Meena H, Solanki K. Effects of Vitrification on Immature and in vitro Matured, Denuded and Cumulus Compact Goat Oocytes and Their Subsequent Fertilization. J Reprod Infertil 2012 Jan;13(1):53-9.
- Combelles CM, Ceyhan ST, Wang H, Racowsky C. Maturation outcomes are improved following Cryoleaf vitrification of immature human oocytes when compared to choline-based slow-freezing. J Assist Reprod Genet 2011 Dec;28(12):1183-92.
- Prentice JR, Anzar M. Cryopreservation of Mammalian oocyte for conservation of animal genetics. Vet Med Int 2010 Sep 21;2011.
- Khosravi-Farsani S, Sobhani A, Amidi F, Mahmoudi R. Mouse oocyte vitrification: the effects of two methods on maturing germinal vesicle breakdown oocytes. J Assist Reprod Genet 2010 May;27(5):233-8.
- Maclellan LJ, Albertini DF, Stokes JE, Carnevale EM. Use of confocal microscopy and intracytoplasmic sperm injection (ICSI) to assess viability of equine oocytes from young and old mares after vitrification. J Assist Reprod Genet 2023 Nov;40(11):2565-2576.
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