Journal of Andrology
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Published-Ahead-of-Print August 1, 2007, DOI:10.2164/jandrol.107.003194

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Factors from damaged sperm affect its DNA integrity and their ability to promote embryo implantation in mice

Miriam Perez-Crespo , Pedro Moreira , Belen Pintado , and Alfonso Gutierrez-Adan *

* To whom correspondence should be addressed. E-mail: agutierr{at}inia.es.

Endogenous nucleases in mouse sperm can be activated by freeze-thawing spermatozoa in media without cryoprotection and cleave spermatozoa DNA. The role of sperm chromatin integrity during intracytoplasmic sperm injection (ICSI) is of critical importance. We have analyzed in B6D2 mouse, the proportion of DNA-fragmented spermatozoa (DFS) produced by incubation in conditioned medium (CM) generated by freeze-thawing sperm in absence of cryoprotection, and the subsequent development, implantation, and offspring obtained after ICSI with incubated spermatozoa. When fresh sperm cells were incubated for 90 min in this CM, a significant increase of DFS was detected by TUNEL assay (27% vs 4.5% in fresh sperm). After ICSI of fresh and incubated spermatozoa, embryos were cultured in vitro either to 2-cell or blastocyst stage, before transferring them into pseudopregnant CD1 females. At day 14, recipients were sacrificed and implantation rates, estimated as number of live foetuses plus resorptions were determined. When ICSI was performed with sperm incubated in CM no effect on fertilization, embryo cleavage, blastocyst rate or blastocyst morphology was detected; however, quality of embryos was affected since total implantation rate decreased significantly (P<0.05) when 2-cells or blastocysts were transferred. Independently of sperm pre-treatment, in vitro culture affected significantly the percentage of live foetuses present on Day 14 of pregnancy. These results demonstrate that there are factors released from fragmented spermatozoa capable of inducing DNA fragmentation in intact sperm cells that may compromise to some extend birth rate after ICSI.


Key words: Assisted reproduction • ICSI • DNA integrity • embryo quality • sperm endonucleases




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