Vitrifying immature equine oocytes impairs their ability to correctly align the chromosomes on the MII spindle
Kaatje D. Ducheyne
A B D , Marilena Rizzo B C , Peter F. Daels A , Tom A. E. Stout B and Marta de Ruijter-Villani B
+ Author Affiliations
- Author Affiliations
A Department of Medical Imaging and Small Animal Orthopaedics, Faculty of Veterinary Medicine, Ghent University, Salisburylaan 133, 9820 Merelbeke, Belgium.
B Department of Equine Sciences, Faculty of Veterinary Medicine, Utrecht University, Yalelaan 112, 3684CM Utrecht, Netherlands.
C Department of Veterinary Sciences, Messina University, Viale Annunziata, 98168 Messina, Italy.
D Corresponding author. Email: k.d.ducheyne@uu.nl
Reproduction, Fertility and Development 31(8) 1330-1338 https://doi.org/10.1071/RD18276
Submitted: 31 July 2018 Accepted: 6 February 2019 Published: 10 April 2019
Abstract
Vitrified–warmed immature equine oocytes are able to complete the first meiotic division, but their subsequent developmental competence is compromised. Therefore, the present study investigated the effects of vitrifying immature horse oocytes on the chromosome and spindle configuration after IVM. Cumulus–oocytes complexes (COCs) were collected and divided into two groups based on mare age (young ≤14 years; old ≥16 years). COCs were then either directly matured in vitro or vitrified and warmed before IVM. Spindle morphology and chromosome alignment within MII stage oocytes were assessed using immunofluorescent staining, confocal microscopy and three-dimensional image analysis. Vitrification reduced the ability of oocytes to reach MII and resulted in ultrastructural changes to the meiotic spindle, including shortening of its long axis, and an increased incidence of chromosomes failing to align properly at the metaphase plate. We hypothesise that aberrant chromosome alignment is an important contributor to the reduced developmental competence of vitrified equine oocytes. Contrary to expectation, oocytes from young mares were more severely affected than oocytes from older mares; we propose that the reduced effect of vitrification on oocytes from older mares is related to pre-existing compromise of spindle assembly checkpoint control mechanisms in these mares.
Additional keywords: chromosome alignment, cryopreservation, horse, meiotic spindle.
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