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Titolo:
Activation of bovine oocytes by specific inhibition of cyclin-dependent kinases
Autore:
Alberio, R; Kubelka, M; Zakhartchenko, V; Hajduch, M; Wolf, E; Motlik, J;
Indirizzi:
Univ Munich, Dept Mol Anim Breeding & Genet, D-85764 Oberschleissheim, Germany Univ Munich Oberschleissheim Germany D-85764 4 Oberschleissheim, Germany Acad Sci Czech Republ, Inst Anim Physiol & Genet, CR-27721 Libechov, CzechRepublic Acad Sci Czech Republ Libechov Czech Republic CR-27721 ov, CzechRepublic Palacky Univ, Fac Med, Dept Pediat, CR-77147 Olomouc, Czech Republic Palacky Univ Olomouc Czech Republic CR-77147 147 Olomouc, Czech Republic
Titolo Testata:
MOLECULAR REPRODUCTION AND DEVELOPMENT
fascicolo: 4, volume: 55, anno: 2000,
pagine: 422 - 432
SICI:
1040-452X(200004)55:4<422:AOBOBS>2.0.ZU;2-R
Fonte:
ISI
Lingua:
ENG
Soggetto:
INTRACYTOPLASMIC SPERM INJECTION; PROTEIN-SYNTHESIS INHIBITION; MATURATION-PROMOTING FACTOR; PARTHENOGENETIC DEVELOPMENT; MAP KINASE; MOUSE EGGS; IN-VITRO; METAPHASE ARREST; CALCIUM SIGNAL; CELL-CYCLE;
Keywords:
bovine; oocyte; parthenogenetic activation; cdk inhibitor;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
69
Recensione:
Indirizzi per estratti:
Indirizzo: Alberio, R Univ Munich, Dept Mol Anim Breeding & Genet, Hackerstr 27, D-85764 Oberschleissheim, Germany Univ Munich Hackerstr 27 Oberschleissheim Germany D-85764 many
Citazione:
R. Alberio et al., "Activation of bovine oocytes by specific inhibition of cyclin-dependent kinases", MOL REPROD, 55(4), 2000, pp. 422-432

Abstract

Activation of bovine oocytes by experimental procedures that closely mimicnormal fertilization and allow to obtain haploid oocytes is essential bothfor intracytoplasmic sperm injection (ICSI) and for nuclear transfer. Therefore, with the goal of producing haploid activated oocytes, this study evaluated whether bohemine, either alone or in combination with ionomycin, is able to activate young matured bovine oocytes. Furthermore, the effect of bohemine on the patterns of DNA synthesis after pronuclear formation as wellas changes in his tone H1 kinase and MAP kinase activities during the process of activation were studied. Our results with bohemine show that the specific inhibition of CDKs in metaphase II bovine oocytes induces parthenogenetic activation in a dose-dependent manner (25, 50, and 100 mu M, respectively), either alone (3%, 30%, and 50%) or in combination with ionomycin (30%, 70%, and 87.5%). A single pronucleus and extrusion of the second polar body was observed (97%) when Ca2+ influx was stimulated in the presence of bohemine, although pronuclear formation without polar body extrusion was observed when bohemine was used alone. Bohemine-activated oocytes started to synthesize DNA in the first hour (37%) after their removal from bohemine-supplemented medium (6-7 hr post-activation; hpa). A high synchrony in the S-phase was registered with more than 85% of parthenotes actively synthesizing DNA 8 hpa. By contrast, DNA synthesis was absent in oocytes cultured for 4,6, and 8 hpa in the presence of bohemine and a low rate was observed by those cultured for 18 hr (30%) in bohemine-supplemented medium. This confirmsthe ability of the inhibitor to arrest the cell cycle in the G1/S boundaryfor at least 8 hr. A drop in histone H1 kinase activity was observed in bohemine-activated oocytes. The activity of MBP kinase decreased later than histone H1 kinase and even 4 hr after inomycin-bohemine treatment at least half of this activity was still detectable. Then, the MBP kinase activity decreased and the lowest level could be seen 6-8 hpa. In summary, our study shows that in vitro matured bovine oocytes can be successfully activated by a synthetic inhibitor of CDKs. This effect can be improved by combination with ionomycin. The targeting of CDKs in the way to activate bovine oocytes can be an approach to improve the efficiency of mammalian oocyte activation. Mel. Re prod. Dev. 55:422-432, 2000. (C) 2000 Wiley-Liss, Inc.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 28/09/20 alle ore 04:11:44