Use of strontium for bovine oocyte activation.
Strontium efficiently activates mouse oocytes, however, there is limited information on its use in cattle. Thus, the objective of this study was to establish a suitable protocol for activating bovine oocyte with strontium. For pronuclear development, the absence of calcium and magnesium in the activation medium (TALP) with 10 and 50 mM strontium (34.4 and 53.1%, respectively) was superior to the complete TALP (6.5 and 19.4%, respectively). In all activation media, better results were observed with 25 and 50 mM strontium (21.9-53.1 and 19.4-53.1%, respectively). Incubation for 4 h promoted similar results in all strontium concentrations. However, strontium at 15, 20, and 25 mM for 6 and 8 h (40.7, 46.7, and 48.3%, and 29.3, 48.3, and 40.7%, respectively) were superior to control (15.5 and 10%, respectively). After in vitro maturation for 26 h, strontium (S; 20 mM in Ca2+- and Mg2+-free TALP for 6 h), ionomycin+strontium (IS), and strontium+ionomycin (SI) (60, 63.3, and 65%, respectively) were similar in pronuclear development and superior to ionomycin (I; 5 microM for 5 min; 36.7%). In treatments S and I, only 1 PN zygotes were observed. In treatment S, most of them had 1 and 2 PB (35.7 and 60.7%, respectively), and in treatment I, 0, 1, and 2 PB (14.3, 57.1, and 28.6%, respectively). Most of the zygotes in treatment IS and SI were 1 PN 2 PB (77.4 and 61.6%, respectively). The number of oocytes with clusters of cortical granules was similar in all treated groups (11-29%). Cortical granule exocytosis in treatment IS (68%) was similar to S (54%) and superior to I, SI, and control (27, 45, and 5.0%, respectively). Cleavage and blastocyst rates were similar for S, I, IS, and SI treatments (61.7-76.7, and 8.3-13.3%, respectively) and the same was observed for ICM, TE, and total cell number, and ICM/total cell ratio (22-25, 64-69, and 86-95, and 0.26-0.27). In conclusion, strontium may be efficiently applied for bovine oocyte activation at 20 mM in Ca2+- and Mg2+-free TALP medium for 6 h.
Méo
Simone Cristina
SC
Faculdade de Ciências Agrárias e Veterinárias, UNESP, Câmpus de Jaboticabal, Via de Acesso Paulo Donato Castellane, s/n 14884-900 Jaboticabal, SP, Brazil. scmeo@fcav.unesp.br
Yamazaki
Walt
W
Leal
Cláudia Lima Verde
CL
de Oliveira
João Ademir
JA
Garcia
Joaquim Mansano
JM
D000818:Animals; D001755:Blastocyst / Q000502:physiology; D002118:Calcium / Q000008:administration & dosage; D002417:Cattle; D002478:Cells, Cultured; D002970:Cleavage Stage, Ovum; D046149:Embryo Culture Techniques / Q000662:veterinary; D005260:Female; D015759:Ionomycin / Q000494:pharmacology; D008274:Magnesium / Q000008:administration & dosage; D009865:Oocytes / Q000187:drug effects* / Q000502:physiology*; D013324:Strontium / Q000008:administration & dosage*
D015759:Ionomycin; D008274:Magnesium; D002118:Calcium; D013324:Strontium