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Effect of vitrification solutions and cooling upon in vitro matured prepubertal ovine oocytes

Succu, Sara and Leoni, Giovanni Giuseppe and Berlinguer, Fiammetta and Madeddu, Manuela and Bebbere, Daniela and Mossa, Francesca and Bogliolo, Luisa and Ledda, Sergio and Naitana, Salvatore (2007) Effect of vitrification solutions and cooling upon in vitro matured prepubertal ovine oocytes. Theriogenology, Vol. 68 (1), p. 107-114. ISSN 0093-691X. Article.

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DOI: 10.1016/j.theriogenology.2007.04.035

Abstract

The vitrification procedure effects on molecular and cytoskeletal components and on developmental ability of in vitro matured prepubertal ovine oocytes were evaluated. MII oocytes were divided into three groups: (1) vitrified in cryoloops (VTR); (2) exposed to vitrification solutions and rehydrated without being plunged into liquid nitrogen (EXP); (3) without further treatment as a control (CTR).
Two hours after treatment, membrane integrity, assessed by propidium iodide/Hoechst staining, was lower in VTR and EXP than in CTR (70.6%, 88.5% and 95.2%, respectively). Cleavage rate after fertilization was statistically different among all groups (21.4%, 45.4% and 82.8% for VTR, EXP and CTR groups respectively; P < 0.01). Blastocyst rate in VTR (0.0%) and EXP (2.8%) groups was lower (P < 0.01) than in CTR (22.8%). Maturation promoting factor activity was lower (P < 0.01) in VTR and EXP groups compared with CTR at both 0 h (82.2%, 83.6% and 100%, respectively) and 2 h (60% and 53.9% and 100%, respectively) after warming.
Immediately after warming VTR and EXP oocytes showed a lower rate of normal spindle and chromosome configuration compared to CTR (59.1%, 48.0% and 83.3%, respectively; P < 0.01). After 2 h of culture in standard conditions the percentage of oocytes with normal spindle and chromosome organization decreased in both VTR and EXP groups compared to CTR (36.4%, 42.8% versus 87.5%, respectively).
In conclusion the exposition to the tested cryoprotectant solution and the vitrification in cryoloops modified cytoskeletal components and alter biochemical pathways that compromise the developmental capacity of prepubertal in vitro matured ovine oocytes.

Item Type:Article
ID Code:4724
Status:Published
Refereed:Yes
Uncontrolled Keywords:Prepubertal, oocyte, vitrification, meiotic spindle, MPF
Subjects:Area 07 - Scienze agrarie e veterinarie > VET/10 Clinica ostetrica e ginecologia veterinaria
Area 07 - Scienze agrarie e veterinarie > VET/02 Fisiologia veterinaria
Area 05 - Scienze biologiche > BIO/10 Biochimica
Divisions:001 Università di Sassari > 01 Dipartimenti > Scienze fisiologiche, biochimiche e cellulari
001 Università di Sassari > 01 Dipartimenti > Patologia e clinica veterinaria
001 Università di Sassari > 01 Dipartimenti > Biologia animale
Publisher:Elsevier
ISSN:0093-691X
Deposited On:22 Oct 2010 13:16

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