Effect of activation with Ca ionophore A23187 and puromycin on the development of human oocytes that failed to fertilize after intracytoplasmic sperm injection

Fertil Steril. 2001 Jul;76(1):148-52. doi: 10.1016/s0015-0282(01)01839-8.


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).

Design: Laboratory examination.

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

Patient(s): Discarded unfertilized oocytes following ICSI.

Intervention(s): 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.

Main outcome measure(s): 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.

Result(s): 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.

Conclusion(s): The sequential treatment of calcium ionophore A23187 and puromycin activates unfertilized oocytes after ICSI. The resultant oocytes with 2PN2PB can cleave.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Calcimycin / pharmacology*
  • Cellular Senescence / drug effects
  • Chromosome Mapping
  • Cleavage Stage, Ovum
  • Diploidy
  • Female
  • Fertilization in Vitro*
  • Humans
  • Ionophores / pharmacology*
  • Male
  • Oocytes / drug effects*
  • Oocytes / physiology*
  • Puromycin / pharmacology*
  • Sperm Injections, Intracytoplasmic*
  • Sperm Motility
  • Spermatozoa / physiology
  • Treatment Failure


  • Ionophores
  • Calcimycin
  • Puromycin