Hostname: page-component-848d4c4894-pftt2 Total loading time: 0 Render date: 2024-05-09T11:48:25.040Z Has data issue: false hasContentIssue false
  • English
  • Français

Ultrastructural Imaging Analysis of the Zona Pellucida Surface in Bovine Oocytes

Part of: Micrographia Collection

Published online by Cambridge University Press:  28 May 2019

Francisco Báez ,
Álvaro A. Camargo  and
Gustavo D.A. Gastal Open the ORCID record for Gustavo D.A. Gastal [Opens in a new window]
Francisco Báez*
Affiliation:
University Center of Tacuarembó, UDELAR, Route 5, Km 386, Tacuarembó, Uruguay
Álvaro A. Camargo
Affiliation:
University Center of Tacuarembó, UDELAR, Route 5, Km 386, Tacuarembó, Uruguay
Gustavo D.A. Gastal
Affiliation:
National Agricultural Research Institute, INIA, Route 50, Km 11, Colonia 70000, Uruguay
*
*Author for correspondence: Francisco Báez, E-mail: francisco.baez@cut.edu.uy
Get access

Abstract

The aims of the present study were to: (i) evaluate the ultrastructural differences in the zona pellucida (ZP) surface between immature and mature bovine oocytes, and (ii) describe a new objective technique to measure the pores in the outer ZP. Intact cumulus–oocyte complexes (COCs) obtained from a local abattoir were immediately fixed (immature group) or submitted to in vitro maturation (IVM) at 38.5 °C for 24 h in a humidified atmosphere of 5% CO2 in air (mature group). Oocytes from both groups were morphologically evaluated via Scanning Electron Microscopy (SEM) and the images were processed in the Fiji/ImageJ software using a new objective methodology through the Trainable Weka Segmentation plugin. The average number of pores in ZP was greater (p < 0.05) in the mature group than the immature group. However, the size and circularity of pores in ZP did not differ (p > 0.05) between groups. In conclusion, it has been shown that the number of pores highlighted the main ultrastructural change in the morphology of the ZP surface of bovine oocytes during the IVM process. We have described an objective method that can be used to evaluate ultrastructural modifications of the ZP surface during oocyte maturation and early embryo development.

Keywords

bovinein vitro maturationoocyteultrastructurezona pellucida
Type
Micrographia
Information
Microscopy and Microanalysis , Volume 25 , Issue 4 , August 2019 , pp. 1032 - 1036
Copyright
Copyright © Microscopy Society of America 2019 

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Arganda-Carreras, I, Kaynig, V, Rueden, C, Eliceiri, KW, Schindelin, J, Cardona, A & Sebastian Seung, H (2017). Trainable Weka Segmentation: A machine learning tool for microscopy pixel classification. Bioinformatics 33(15), 2424–2426.Google Scholar
Choi, BH, Bang, JI, Jin, JI, Kim, SS, Jo, HT, Deb, GK, Ghanem, N, Cho, KW & Kong, IK (2013). Coculturing cumulus oocyte complexes with denuded oocytes alters zona pellucida ultrastructure in in vitro matured bovine oocytes. Theriogenology 80(9), 1117–1123.Google Scholar
Dudkiewicz, A & Williams, W (1977). Fine structural observations of the mammalian zona pellucida by scanning electron microscopy. Scanning Electron Microsc 2, 317–324.Google Scholar
Dunbar, BS, Avery, S, Lee, V, Prasad, S, Schwahn, D, Schwoebel, E, Skinner, S & Wilkins, B (1994). The mammalian zona pellucida: Its biochemistry, immunochemistry, molecular biology, and developmental expression. Reprod Fertil Dev 6(3), 331–347.Google Scholar
Epifano, O & Dean, J (1994). Biology and structure of the zona pellucida: A target for immunocontraception. Reprod Fertil Dev 6(3), 319–330.Google Scholar
Familiari, G, Relucenti, M, Heyn, R, Micara, G & Correr, S (2006). Three-dimensional structure of the zona pellucida at ovulation. Microsc Res Tech 69(6), 415–426.Google Scholar
Held, E, Mertens, EM, Mohammadi-Sangcheshmeh, A, Salilew-Wondim, D, Besenfelder, U, Havlicek, V, Herrler, A, Tesfaye, D, Schellander, K & Holker, M (2012). Zona pellucida birefringence correlates with developmental capacity of bovine oocytes classified by maturational environment, COC morphology and G6PDH activity. Reprod Fertil Dev 24(4), 568–579.Google Scholar
Hewitt, DA, Watson, PF & England, GCW (1998). Nuclear staining and culture requirements for in vitro maturation of domestic bitch oocytes. Theriogenology 49(6), 1083–1101.Google Scholar
Magerkurth, C, Topfer-Petersen, E, Schwartz, P & Michelmann, HW (1999). Scanning electron microscopy analysis of the human zona pellucida: Influence of maturity and fertilization on morphology and sperm binding pattern. Hum Reprod 14(4), 1057–1066.Google Scholar
Moreira da Silva, F & Metelo, R (2005). Relation between physical properties of the zona pellucida and viability of bovine embryos after slow-freezing and vitrification. Reprod Domest Anim 40(3), 205–209.Google Scholar
Riddell, KP, Stringfellow, DA, Gray, BW, Riddell, MG, Wright, JC & Galik, PK (1993). Structural and viral association comparisons of bovine zonae pellucidae from follicular oocytes, Day-7 embryos and Day-7 degenerated ova. Theriogenology 40(6), 1281–1291.Google Scholar
Santos, P, Chaveiro, A, Simoes, N & Moreira da Silva, F (2008). Bovine oocyte quality in relation to ultrastructural characteristics of zona pellucida, polyspermic penetration and developmental competence. Reprod Domest Anim 43(6), 685–689.Google Scholar
Schindelin, J, Arganda-Carreras, I, Frise, E, Kaynig, V, Longair, M, Pietzsch, T, Preibisch, S, Rueden, C, Saalfeld, S, Schmid, B, Tinevez, JY, White, DJ, Hartenstein, V, Eliceiri, K, Tomancak, P & Cardona, A (2012). Fiji: An open-source platform for biological-image analysis. Nat Methods 9(7), 676–682.Google Scholar
Suzuki, H, Yang, X & Foote, RH (1994). Surface alterations of the bovine oocyte and its investments during and after maturation and fertilization in vitro. Mol Reprod Dev 38(4), 421–430.Google Scholar
Vanroose, G, Nauwynck, H, Soom, AV, Ysebaert, MT, Charlier, G, Oostveldt, PV & de Kruif, A (2000). Structural aspects of the zona pellucida of in vitro-produced bovine embryos: A scanning electron and confocal laser scanning microscopic study. Biol Reprod 62(2), 463–469.Google Scholar
Wassarman, PM (1990). Regulation of mammalian fertilization by zona pellucida glycoproteins. J Reprod Fertil 42, 79–87.Google Scholar
Zhao, H, Ji, Q, Zhao, G, Song, Z, Du, B, Nie, Y, Chen, Y & Cong, P (2014). Damage of zona pellucida reduces the developmental potential and quality of porcine circovirus type 2-infected oocytes after parthenogenetic activation. Theriogenology 82(6), 790–799.Google Scholar