Investigating Spindle Assembly in Oocytes

The function of the two kinases Mps1 and AurkC was investigated using two inhibitory drugs, namely Mps1i or compound 5 (kindly supplied by Prof. Geert Kops Hubrecht Institute, Utrecht, the Netherlands) (Kwiatkowski et al., 2010 (link), p. 359–368) and AURKi or ZM447439 (CAS 331771, Sigma-Aldrich Chemical Co., St. Louis, Missouri, United States). In order to evaluate the function of Mps1 and AurkC on spindle assembly, mature oocytes displaying a polar body were first incubated for 10 min at 38°C with 5% CO₂ in IVM medium containing 20 μM Nocodazole (M1404, Sigma-Aldrich Chemical Co., St. Louis, Missouri, United States) to depolymerize the MII-spindle. Following Nocodazole washout, the oocytes were left to re-form their spindle for 120 min in IVM medium with different concentrations of either MPS1i (0, 200, or 500 nM) or AURKi (0, 5, or 10 μM). The oocytes in the control group were not treated with Nocodazole or inhibitors.

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Rizzo M., Stout T.A., Cristarella S., Quartuccio M., Kops G.J, & De Ruijter-Villani M. (2021). Compromised MPS1 Activity Induces Multipolar Spindle Formation in Oocytes From Aged Mares: Establishing the Horse as a Natural Animal Model to Study Age-Induced Oocyte Meiotic Spindle Instability. Frontiers in Cell and Developmental Biology, 9, 657366.

Publication 2021

ZM447439 Nocodazole

Drugs Inhibitors Inhibitory Kinases Mps1 Nocodazole Oocytes Polar body Zm447439

Corresponding Organization : Utrecht University

Other organizations : University of Messina, Oncode Institute, University Medical Center Utrecht

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Variable analysis

independent variables

Mps1i (0, 200, or 500 nM)
AURKi (0, 5, or 10 μM)

dependent variables

Spindle assembly

control variables

Mature oocytes displaying a polar body
Incubation for 10 min at 38°C with 5% CO2 in IVM medium containing 20 μM Nocodazole to depolymerize the MII-spindle
Re-formation of spindle for 120 min in IVM medium

controls

Positive control: Oocytes not treated with nocodazole or inhibitors

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