Oda S, Deguchi R, Mohri T, Shikano T, Nakanishi S, Miyazaki S
Department of Physiology, Tokyo Women's Medical University School of Medicine, Shinjuku-ku, Tokyo, 162-8666, Japan.
Dev Biol. 1999 May 1;209(1):172-85. doi: 10.1006/dbio.1999.9233.
Hamster sperm extract (SE) possessing Ca2+ oscillation-inducing activity was microinjected into the peripheral or central region of mouse eggs, and the first increase in intracellular Ca2+ concentration ([Ca2+]i), together with the spread of fluorescence-labeled SE in the ooplasm, was investigated by imaging with confocal microscopy. Injection into the periphery always induced a Ca2+ wave that started from the injection site after a delay of 5 to 30 s depending on the concentration of SE. The diluted SE caused a wave of two-step [Ca2+]i rises, which was always observed at fertilization. Injection into the center could induce a radial Ca2+ wave with relatively high dose of SE, but lower dose of SE caused a [Ca2+]i rise after a longer delay which was initiated synchronously over the ooplasm or was preceded in a peripheral area. Injection of diluted SE remarkably prolonged the delay time and reduced the rate of [Ca2+]i rise. The critical concentration of SE needed to induce [Ca2+]i rise was significantly lower in the periphery. These results indicate that the sensitivity to SE is higher in the cortex. SE-induced [Ca2+]i rises were blocked by an antibody against the type 1 inositol 1,4,5-trisphosphate receptor (InsP3R). The cortex was substantially more sensitive to injected InsP3 induction of Ca2+ release than the center. It is suggested that the cortex of mouse eggs may involve a functionally specialized organization of InsP3Rs and Ca2+ pools in which a cytosolic sperm factor(s) could act upon sperm-egg fusion to cause Ca2+ release, leading to the Ca2+ wave at fertilization.
将具有诱导钙离子振荡活性的仓鼠精子提取物(SE)显微注射到小鼠卵母细胞的外周或中央区域,并用共聚焦显微镜成像研究细胞内钙离子浓度([Ca2+]i)的首次升高以及荧光标记的SE在卵质中的扩散情况。注射到外周区域总是会诱导出钙离子波,延迟5到30秒后从注射部位开始,延迟时间取决于SE的浓度。稀释后的SE会引起两步[Ca2+]i升高的波,这在受精时总是能观察到。注射到中央区域,相对高剂量的SE可诱导出径向钙离子波,但低剂量的SE会在更长的延迟后引起[Ca2+]i升高,这种升高在整个卵质中同步开始或在外周区域先出现。注射稀释后的SE会显著延长延迟时间并降低[Ca2+]i升高的速率。在外周区域诱导[Ca2+]i升高所需的SE临界浓度显著更低。这些结果表明皮质对SE的敏感性更高。SE诱导的[Ca2+]i升高被抗1型肌醇1,4,5-三磷酸受体(InsP3R)的抗体阻断。皮质对注射的InsP3诱导的钙离子释放比中央区域更敏感。提示小鼠卵母细胞的皮质可能涉及InsP3Rs和钙离子池的功能特化组织,其中一种胞质精子因子可作用于精卵融合以引起钙离子释放,导致受精时的钙离子波。