Catalase in manipulation buffer enhances the developmental competence of DNA-injected embryos.
To improve the efficiency of transgenesis, we investigated the effects of a radical scavenger during microinjection on the development to blastocysts or pups of mouse pronuclear embryos, microinjected with the enhanced green fluorescent protein (EGFP) transgene. When embryos were microinjected in medium containing 0-1,000 units/ml catalase, the developmental rate to blastocysts was significantly higher (P<0.01) in 100-units/ml catalase (81%) than those in 0 and 1,000 units/ml (56 and 65%). To investigate the ontogenetic ability of DNA-injected embryos, EGFP-injected embryos manipulated under 0 or 100 units/ml catalase were transferred separately to recipient mice. The proportion of fetuses derived from EGFP-injected embryos manipulated under 100 units/ml catalase (29%) was significantly higher (P<0.05) than that manipulated under 0 units/ml catalase (19%). Furthermore, the numbers of transgenic pups were 17 in 100 units/ml catalase and 14 in 0 units/ml catalase. The results of the present study indicate that scavenging reactive oxygen species during in vitro micromanipulation is beneficial for the development of DNA-injected embryos.
Hashimoto
Shu
S
The YS Institute Inc., Tochigi 321-0973, Japan. hashimoto@ivfnamba.com
Kuramochi
Takashi
T
Aoyagi
Kazuki
K
Takahashi
Ri-ichi
R
Ueda
Masatsugu
M
D000818:Animals; D001755:Blastocyst / Q000378:metabolism; D002374:Catalase / Q000378:metabolism*; D002467:Cell Nucleus / Q000378:metabolism; D004247:DNA / Q000378:metabolism*; D046149:Embryo Culture Techniques / Q000379:methods*; D004622:Embryo, Mammalian / Q000378:metabolism; D005260:Female; D018014:Gene Transfer Techniques; D005821:Genetic Techniques; D005838:Genotype; D049452:Green Fluorescent Proteins / Q000378:metabolism; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D017382:Reactive Oxygen Species; D013094:Spermatozoa / Q000378:metabolism; D019076:Transgenes
D017382:Reactive Oxygen Species; D049452:Green Fluorescent Proteins; D004247:DNA; D002374:Catalase