The effects of microtubule stabilizing and recovery agents on vitrified bovine oocytes.

Emily Girka, Lauren Gatenby, Emilio J Gutierrez, Kenneth R Bondioli

Theriogenology 2022 Apr 01

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Oocyte vitrification, while beneficial for research and species conservation applications, has limited success due to cryoinjury to the meiotic spindle. This study aimed to improve meiotic spindle recovery in vitrified bovine oocytes by investigating the effects of treatment with either a microtubule stabilizing agent, or a microtubule recovery agent. In the first two experiments, either paclitaxel or epothilone B were used to treat bovine oocytes before vitrification. Both compounds have microtubule stabilizing properties and are known antimitotic compounds used to disrupt microtubule dynamics in rapidly proliferating cancer cells. Paclitaxel treatment at 2.0 μM significantly increased the proportion of oocytes with normal microtubule distribution and chromosome arrangement after warming. Treatment with 1.0 μM had no effect and 0.5 μM had a negative effect on meiotic spindle recovery. Epothilone B treatment at all concentrations significantly increased the proportion of oocytes with meiotic spindle disruption and abnormally dispersed chromosomes. In the second set of experiments, Rho-associated coiled-coil kinase inhibition and glutathione accumulation were investigated as recovery treatments after vitrification. Oocytes were incubated with either Y-27632 or combinations of cysteine and cysteamine for 4 h after warming. Treatment with 5 μM and 10 μM of Y-27632 to inhibit rho-associated coiled-coil kinase activity significantly increased the proportion of vitrified oocytes with normal microtubule distribution and chromosome arrangement. When oocytes were incubated with 20 μM of Y-27632 there was no effect on spindle recovery. Incubation with 100 μM of cysteamine also had no effect on spindle recovery while 0.6 mM of cysteine and both 0.6 mM of cysteine and 100 μM of cysteamine significantly increased oocytes with normal microtubule distribution and chromosome arrangement. Copyright © 2022 Elsevier Inc. All rights reserved.

Citation

Emily Girka, Lauren Gatenby, Emilio J Gutierrez, Kenneth R Bondioli. The effects of microtubule stabilizing and recovery agents on vitrified bovine oocytes. Theriogenology. 2022 Apr 01;182:9-16