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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
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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 

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