Effects of heat shock on in vitro development and intracellular oxidative state of bovine preimplantation embryos
Effects of heat shock on in vitro development and intracellular oxidative state of bovine preimplantation embryos
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DOI：
10.1002/mrd.20014
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发表时间：
2004-01-01
影响因子：
2.5
通讯作者：
Takahashi, M
中科院分区：
文献类型：
--
作者：
Sakatani, M;Kobayashi, SI;Takahashi, M
研究了热休克对牛胚胎发育能力和细胞内氧化状态的影响。体外受精后，胚胎分别在第0、2、4和6天在41 ℃下热休克6小时。在第2天，卵裂率在所有组中没有显著差异。然而，与对照组（37.5+/-4.0%）、第4天（40.0+/-7.4%）和第6天（38.1+/-2.0%）相比，暴露于热休克后第0天（18.8+/-4.3%）和第2天（23.6+/-3.7%）发育至囊胚期的胚胎百分比显著降低。此外，与对照组（143.2+/-9.4）相比，热休克第0天（107.5+/-6.6）和第2天（112.8+/-5.7）的囊胚总细胞数显著减少。为了评价热休克后细胞内氧化状态，将热休克后0、2、4、6 d的胚胎与2 '，7'-二氯二氢荧光素二乙酸酯（DCHFDA）共同孵育，在荧光显微镜下检测活性氧（ROS）氧化DCHFDA的荧光。当胚胎暴露于热休克0和2天的荧光强度显着增加。然而，在第4天和第6天的热休克没有增加荧光强度。这些结果表明：（1）早期胚胎的热休克导致囊胚发育和细胞增殖的减少;（2）热休克导致的发育减少可能与细胞内氧化应激的增加有关。
We investigated the effects of heat shock on developmental competence of bovine embryos and intracellular oxidative state. After in vitro fertilization, embryos were exposed to heat shock at 41degreesC for 6 hr on days 0, 2, 4, and 6, respectively. On day 2, cleavage rate was not significantly different in all groups. However, the percentage of embryos developing to blastocyst stage after exposure to heat shock on day 0 (18.8+/-4.3%) and day 2 (23.6+/-3.7%) were significantly decreased compared with control (37.5+/-4.0%), day 4 (40.0+/-7.4%), and day 6 (38.1+/-2.0%). In addition, the total cell number of blastocysts was significantly decreased by heat shock on day 0 (107.5+/-6.6) and day 2 (112.8+/-5.7) compared with the control (143.2+/-9.4). To evaluate intracellular oxidative state by heat shock, embryos exposed to heat shock on days 0, 2, 4, and 6 were incubated with 2',7'-dichlorodihydrofluorescein diacetate (DCHFDA) and fluorescence of oxidized DCHFDA by reactive oxygen species (ROS) was detected under fluorescent microscope. The intensity of fluorescence was significantly increased when embryos were exposed to heat shock on days 0 and 2. However, heat shock on day 4 and day 6 did not increase the fluorescence intensity. These results indicate that (1) heat shock to earlier stage embryos causes a decrease in development to blastocysts and cell proliferation and (2) the decrease in development by heat shock could be involved in an increase of intracellular oxidative stress.