In vitro viability and ultrastructural changes in bovine oocytes treated with a vitrification solution.

Abstract:

:Abattoir-derived oocytes were exposed to a concentrated cryoprotectant solution (DAP213: 2 M DMSO, 1 M acetamide, 3 M propanediol, and 10% FCS in TCM199) for 1.5 or 5 min at the germinal vesicle (GV) stage or after maturation in vitro (IVM). Their viability was assessed by in vitro fertilization (IVF) and culture (IVC) to blastocysts. To investigate the effect of DAP213 on the ultrastructure, GV and IVM oocytes were processed for transmission electron microscopy (TEM) before (control) or after exposure to the cryoprotectant. DAP213 induced profound ultrastructural modifications to the microvilli and mitochondria, resulted in large vesicle formation, and, most significantly, caused the premature release of the cortical granules (CG). In IVM oocytes exposed to the cryoprotectant for 5 min, exocytosis of CG into the perivitelline space was common and the IVF rate was reduced (P < .05). After exposure for 5 min, GV oocytes displayed clusters of CG comparable to controls, but after IVM-IVF, polyspermy rate was increased (P < .05). Furthermore, treated GV oocytes showed a reduced rate of cleavage and blastocyst formation and an increased percentage of oocytes exhibiting alterations in organelles, whereas the viability and ultrastructure of IVM oocytes treated for 1.5 min was not different from controls. These observations demonstrate that 1) cortical granule kinetics is one of the key elements controlling fertilizability of bovine oocytes treated with cryoprotectant, and 2) GV oocytes are more sensitive to the cryoprotectant than those that have already been matured in vitro.

journal_name

Mol Reprod Dev

authors

Fuku EJ,Liu J,Downey BR

doi

10.1002/mrd.1080400206

subject

Has Abstract

pub_date

1995-02-01 00:00:00

pages

177-85

issue

2

eissn

1040-452X

issn

1098-2795

journal_volume

40

pub_type

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