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Key Role for Cyclin-Dependent Kinases in the First and Second Meiotic Divisions of Rat Spermatocytes¹

INSERM U418/INRA UMR 1245,³ Hôpital Debrousse, 69322 Lyon, cedex 05, France Laboratoire de Biologie Moléculaire de la Cellule,⁴ CNRS UMR 5161, Ecole Normale Supérieure de Lyon, 69364 Lyon cedex 07, France

In all systems examined so far, the G2/M phase transition is controlled by the M-phase promoting factor (MPF), a complex of cdc2 (CDK1) and cyclin B1. Histone H1 kinase activity and MPF components are present in pachytene spermatocytes (PS). However, it has not been demonstrated yet that direct inhibition of MPF activity prevents the G2/M transition in these cells. When roscovitine, a potent inhibitor of CDK1, CDK2, and CDK5 activities, was added to cocultures of PS with Sertoli cells, the number of both secondary spermatocytes and round spermatids formed were lower than in control cultures, despite similar cell viability. This effect of roscovitine was reversible, did not involve the Sertoli cells, and was dependent on the concentration of the inhibitor. Roscovitine did not modify the amount of MPF in these germ cells but inhibited the CDK1- or CDK2-associated histone H1 kinase activity of PS. Hence a functional relationship between cyclin-dependent kinase activity and the spontaneous processing of the first meiotic division and, for the first time, of the second meiotic division of male germ cells is shown.

¹ Supported by the Institut National de la Santé et de la Recherche Médicale, the Institut National de la Recherche Agronomique, the Centre National pour la Recherche Scientifique, the Ministry of Research and Technology, and the Université Claude Bernard Lyon I.

² Correspondence: Murielle Godet, INSERM U418/INRA UMR 1245, Hôpital Debrousse, 29, Rue Sur Bouvier, 69322 Lyon cedex 05, France. FAX: 33 4 78256168; godet{at}lyon.inserm.fr

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