Effects of heat shock on in vitro development and intracellular oxidative state of bovine preimplantation embryos.
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 41 degrees C 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. Mol. Reprod. Dev. 67: 77-82, 2004.
Sakatani
Miki
M
Department of Animal and Grassland Research, National Agricultural Research Center for Kyushu Okinawa Region, Nishigoshi, Kikuchi, Kumamoto, Japan. msaka@affrc.go.jp
Kobayashi
Shu-Ichi
S
Takahashi
Masashi
M
D000818:Animals; D001755:Blastocyst / Q000378:metabolism / Q000502:physiology*; D002417:Cattle; D005260:Female; D005452:Fluoresceins / Q000378:metabolism; D006358:Hot Temperature / Q000009:adverse effects*; D008297:Male; D010084:Oxidation-Reduction; D018384:Oxidative Stress; D011247:Pregnancy; D017382:Reactive Oxygen Species / Q000378:metabolism
C110400:2',7'-dichlorodihydrofluorescein diacetate; D005452:Fluoresceins; D017382:Reactive Oxygen Species