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Differential effects of 6-DMAP, olomoucine and roscovitine on Xenopus oocytes and eggs

  • Stéphane Flament (a1), Jean-François Bodart (a1), Marc Bertout (a1), Edith Browaeys (a1), Arlette Rousseau (a1) and Jean-Pierre Vilain (a1)...

Abstract

The effects of the new cyclin-dependent kinase inhibitors, roscovitine and olomoucine, on oocytes and eggs of Xenopus laevis were investigated and compared with those of 6-dimethylamino purine (6-DMAP). The inhibitory properties of 6-DMAP, olomoucine and roscovitine towards p34cdc2-cyclin B isolated from Xenopus eggs revealed K-IC50 values of 300, 40 and 10 μM respectively. The three compounds inhibited progesterone-induced maturation with M-IC50 values of 200, 100 and 20 μM. These values were consistent with the K-IC50 values but the ratio M-IC50/K-IC50 was higher for roscovitine and olomoucine than for 6-DMAP. The disappearance of spindle and condensed chromosomes without pronucleus formation was observed when 1 mM 6-DMAP was applied for 4 h at germinal vesicle breakdown or at metaphase II, whereas no effect was observed using 1 mM olomoucine or 50 μM roscovitine. Changes in the electrophoretic mobility of p34cdc2 and erk2 were observed only in homogenates of matured oocytes or eggs exposed for 4 h to 1 mM 6-DMAP. When the drugs were microinjected into matured oocytes, olomoucine (100 μM) and roscovitine (50 μM) induced pronucleus formation more efficiently than did 6-DMAP (100 μM). Taken together, these results demonstrate that Xenopus oocytes possess a lower permeability to olomoucine and roscovitine and that these new compounds are suitable for in vivo studies after germinal vesicle breakdown provided they are microinjected.

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Centre de Biologie Cellulaire, Laboratoire de Biologie du Développement, UPRES EA 1033, Université de Lille 1, SN3, F-59655 Villeneuve d'Ascq cedex, France. Tel: +33 03 20 33 61 16. Fax: +33 03 20 43 40 38.

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Differential effects of 6-DMAP, olomoucine and roscovitine on Xenopus oocytes and eggs

  • Stéphane Flament (a1), Jean-François Bodart (a1), Marc Bertout (a1), Edith Browaeys (a1), Arlette Rousseau (a1) and Jean-Pierre Vilain (a1)...

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