Bovine oocytes treated prior to in vitro maturation with a combination of butyrolactone I and roscovitine at low doses maintain a normal developmental capacity.
Butyrolactone I (BL-I) and Roscovitine (ROS), two specific and potent inhibitors of M-phase promoting factor (MPF) kinase activity, were used to block germinal vesicle breakdown (GVBD) of cattle oocytes. A concentration 6.25 microM BL-I and 12.5 microM ROS blocked over 93.3 +/- 2.5% of oocytes in germinal vesicle (GV) stage during a 24-hr culture period. Following a second 24-hr culture step in maturation medium (IVM) almost all (91.5 +/- 3.0%) inhibited oocytes resumed meiosis and reached the metaphase II (MII) stage. The MII kinetics was different for inhibited and control oocytes. Fifty percent MII was reached at 13-14 hr in BL-I + ROS treated oocytes, compared to 18 hr in control oocytes. Therefore, control oocytes were fertilised (IVF) after 22 hr IVM and inhibited oocytes after 16 or 22 hr IVM. After IVF, percentage of grade 1 freezable embryos on day 7 (D + 7) as well as percentage of blastocyst formation on D + 8 in the group of BL-I + ROS treated oocytes fertilised after 16 hr IVM were higher (P < 0.05) compared with the other experimental group fertilised after 22 hr IVM but not different in comparison with the control. Survival to freezing and thawing of grade 1 embryos frozen on D + 7 was employed as viability criteria and was similar in all groups. Thus, the presence of BL-I + ROS in the prematuration medium of bovine oocytes determines a reversible meiotic block, without compromising their subsequent developmental competence.
Ponderato
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Laboratorio di Tecnologie della Riproduzione (LTR-CIZ), Via Porcellasco 7/f, 26100 Cremona, Italy.
Lagutina
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Crotti
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Turini
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Galli
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C
Lazzari
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D015107:4-Butyrolactone / Q000008:administration & dosage / Q000031:analogs & derivatives* / Q000494:pharmacology*; D000818:Animals; D002417:Cattle; D002467:Cell Nucleus / Q000187:drug effects / Q000378:metabolism; D004357:Drug Synergism; D004622:Embryo, Mammalian / Q000166:cytology / Q000187:drug effects / Q000196:embryology; D005314:Embryonic and Fetal Development / Q000187:drug effects; D005260:Female; D005615:Freezing; D066298:In Vitro Techniques; D008540:Meiosis / Q000187:drug effects; D009865:Oocytes / Q000166:cytology / Q000187:drug effects* / Q000254:growth & development*; D011687:Purines / Q000008:administration & dosage / Q000494:pharmacology*; D000077546:Roscovitine; D015996:Survival Rate; D013997:Time Factors
D011687:Purines; D000077546:Roscovitine; C082868:butyrolactone I; D015107:4-Butyrolactone