Effect of activation with Ca ionophore A23187 and puromycin on the development of human oocytes that failed to fertilize after intracytoplasmic sperm injection.
OBJECTIVE
To determine the effect of sequential oocyte activation with calcium ionophore A23187 (A23187) and puromycin on oocytes that were unfertilized after intracytoplasmic sperm injection (ICSI).


METHODS
Laboratory examination.


METHODS
The in vitro fertilization/embryo transfer unit in Tokushima University Hospital.


METHODS
Discarded unfertilized oocytes following ICSI.


METHODS
All 172 oocytes that showed no evidence of normal fertilization 18 hours after ICSI were exposed to A23187 (5 microM) for 5 minutes and consequently were treated with puromycin (10 microg/mL) for 5 hours.


METHODS
The activation rate, proportion of oocytes that showed two pronuclei with the second polar body (2PN2PB), and cleavage rate were calculated. Chromosomal analysis of the oocytes with 2PN2PB was also performed.


RESULTS
The treatment activated 146 out of 172 oocytes (84.9%) and 30.1% of the activated oocytes showed 2PN2PB. Sixteen of 25 oocytes with 2PN2PB (64.0%) cleaved. There was no difference in the activation rate, proportion of activated oocytes with 2PN2PB, or cleavage rate between oocytes that were injected with a motile spermatozoon and those injected with an immotile spermatozoon. Chromosomal analysis showed a normal diploid set of chromosomes (n = 46) in four of five analyzable oocytes.


CONCLUSIONS
The sequential treatment of calcium ionophore A23187 and puromycin activates unfertilized oocytes after ICSI. The resultant oocytes with 2PN2PB can cleave.
Nakagawa
K
K
Department of Obstetrics and Gynecology, School of Medicine, The University of Tokushima, Tokushima, Japan.
Yamano
S
S
Moride
N
N
Yamashita
M
M
Yoshizawa
M
M
Aono
T
T
D000001:Calcimycin / Q000494:pharmacology*; D016922:Cellular Senescence / Q000187:drug effects; D002874:Chromosome Mapping; D002970:Cleavage Stage, Ovum; D004171:Diploidy; D005260:Female; D005307:Fertilization in Vitro; D006801:Humans; D007476:Ionophores / Q000494:pharmacology*; D008297:Male; D009865:Oocytes / Q000187:drug effects* / Q000502:physiology*; D011691:Puromycin / Q000494:pharmacology*; D020554:Sperm Injections, Intracytoplasmic; D013081:Sperm Motility; D013094:Spermatozoa / Q000502:physiology; D017211:Treatment Failure
D007476:Ionophores; D000001:Calcimycin; D011691:Puromycin