INTRACELLULAR CA2+ RESPONSE OF RABBIT OOCYTES TO ELECTRICAL-STIMULATION

被引:82
作者
FISSORE, RA [1 ]
ROBL, JM [1 ]
机构
[1] UNIV MASSACHUSETTS, DEPT VET & ANIM SCI, AMHERST, MA 01003 USA
关键词
ACTIVATION; CALCIUM; FURA-2; ELECTRICAL PULSE;
D O I
10.1002/mrd.1080320103
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Electrical stimulation is known to cause activation in mammalian oocytes, possibly by eliciting an elevation in intracellular calcium (Ca2+). This study reports intracellular Ca2+ concentrations in mature rabbit oocytes using the Ca2+ indicator fura-2. Calcium levels were determined prior to, during, and after the administration of an electrical pulse (3.6 kV/cm for 60-mu-sec). Baseline Ca2+ levels ranged from 30 to 90 nM. The intracellular Ca2+ transient evoked by a pulse, peaked at 11 sec, was highly variable in amplitude (40-300 nM) and returned to prepulse levels within 300 sec. Electrically stimulated oocytes did not exhibit repetitive Ca2+ transients. The size of the cytoplasmic Ca2+ rise was influenced by the duration of the pulse, the field strength and the concentrations of external Ca2+ (p < 0.05). Oocytes electrically stimulated in the presence of 100-mu-M CaCl2, which evoked Ca2+ transients with a mean magnitude of 120 nM, activated at a higher rate (P < 0.05) than oocytes stimulated in the presence of either higher or lower levels of external Ca2+. Although oocytes electrically shocked at 16-18 hr after administration of human chorionic gonadotropin (hphCG) activated at a lower rate than oocytes stimulated at 22-24 hphCG (P < 0.05), their intracellular Ca2+ response to the pulse was similar (P < 0.05). These results indicate that electrical pulse parameters and extracellular Ca2+ concentrations can be used to modulate intracellular Ca2+ levels and optimize oocyte activation rates. Furthermore, the data suggest that as the oocyte ages it becomes more responsive to a given intracellular Ca2+ elevation.
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页码:9 / 16
页数:8
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