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  • Title: Apoptosis in human unfertilized oocytes after intracytoplasmic sperm injection.
    Author: Bosco L, Ruvolo G, Morici G, Manno M, Cittadini E, Roccheri MC.
    Journal: Fertil Steril; 2005 Nov; 84(5):1417-23. PubMed ID: 16275238.
    Abstract:
    OBJECTIVE: To investigate the presence of programmed cell death in unfertilized oocytes after intracytoplasmic sperm injection (ICSI), assuming that previous apoptotic events could be correlated with the fertilization failure. DESIGN: Comparison of the rate of DNA fragmentation in human oocytes at different stages of maturation soon after pick-up (control) and in unfertilized oocytes after ICSI treatment. SETTING: In vitro fertilization (IVF) laboratory with extensive ICSI experience. PATIENT(S): Sixty-three patients undergoing assisted fertilization by ICSI. INTERVENTION(S): Terminal deoxynucleotidyl transferase-mediated digoxigenin-dUTP nick-end labeling (TUNEL) assay and anticaspase-3 cleaved immunoassay to detect apoptosis in control and ICSI-treated oocytes. MAIN OUTCOME MEASURE(S): Differences in the percentage of oocytes demonstrating DNA fragmentation between control oocytes and unfertilized ICSI treated oocytes at different stages of maturation. RESULT(S): The DNA fragmentation, by TUNEL assay, appeared in all the immature control oocytes, but only 37% of mature oocytes showed DNA fragmentation. This DNA fragmentation was observed in 88.8% of the oocytes unfertilized after ICSI; furthermore, DNA fragmentation appeared as well in the sperm injected into the cytoplasm. CONCLUSION(S): The study has shown DNA fragmentation in human oocytes unfertilized after ICSI. The evidence is confirmed as well in control oocytes, free from in vitro culture or manipulation stress. Caspase-3 immunoassay suggests the presence of apoptosis. The high percentage of oocytes demonstrating DNA fragmentation in the unfertilized oocytes could be correlated with fertilization failure.
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