Swann K
M.R.C. Experimental Embryology and Teratology Unit, St. George's Hospital Medical School, London, U.K.
Biochem J. 1992 Oct 1;287 ( Pt 1)(Pt 1):79-84. doi: 10.1042/bj2870079.
Relative intracellular free Ca2+ concentrations ([Ca2+]i) were monitored in mature unfertilized mouse eggs by measuring fluorescence of intracellular fluo3. A number of different agents were found to cause sustained repetitive transient [Ca2+]i oscillations. These were microinjection of a cytosolic sperm factor, sustained injection of Ins-(1,4,5)P1, or extracellular addition of the thiol reagent thimerosal. Stimulating G-protein activity by injection of guanosine 5'-[gamma-thio]triphosphate plus application of carbachol also caused [Ca2+]i oscillations, but less reliably than other stimuli. A role for Ca(2+)-induced Ca2+ release and a ryanodine-sensitive Ca2+ channel in mouse eggs was suggested by the finding that microinjection, or external addition, of ryanodine also caused [Ca2+]i increases. Furthermore, ryanodine, along with thimerosal, increased the sensitivity of eggs to Ca(2+)-induced [Ca2+]i oscillations. When ryanodine was added to eggs oscillating in response to the sperm factor, InsP3 or thimerosal, it caused a decrease in amplitude of oscillations and eventually a block of [Ca2+]i oscillations associated with a sustained elevation of [Ca2+]i. These data suggest that a ryanodine-sensitive Ca(2+)-release mechanism exists in mouse eggs and that a ryanodine-sensitive Ca2+ store plays a role in generating intracellular [Ca2+]i oscillations.
通过测量细胞内荧光素3的荧光强度,监测成熟未受精小鼠卵中的相对细胞内游离钙离子浓度([Ca2+]i)。发现多种不同试剂可引起持续性重复短暂的[Ca2+]i振荡。这些试剂包括显微注射一种胞质精子因子、持续注射Ins-(1,4,5)P1或细胞外添加硫柳汞。注射鸟苷5'-[γ-硫代]三磷酸并应用卡巴胆碱刺激G蛋白活性也会引起[Ca2+]i振荡,但不如其他刺激可靠。发现显微注射或外部添加ryanodine也会导致[Ca2+]i增加,这提示了Ca(2+)诱导的Ca2+释放和ryanodine敏感的Ca2+通道在小鼠卵中的作用。此外,ryanodine与硫柳汞一起增加了卵对Ca(2+)诱导的[Ca2+]i振荡的敏感性。当ryanodine添加到因精子因子、InsP3或硫柳汞而振荡的卵中时,它会导致振荡幅度减小,最终阻断与[Ca2+]i持续升高相关的[Ca2+]i振荡。这些数据表明,小鼠卵中存在ryanodine敏感的Ca(2+)释放机制,并且ryanodine敏感的Ca2+储存库在产生细胞内[Ca2+]i振荡中起作用。