Early pre-pubertal exposure to low-dose oral di(2-ethylhexyl) phthalate does not affect sperm plasma membrane stability, acrosomal integrity or chromatin structure in the post-pubertal boar

摘要

This study aimed to determine whether pre-pubertal exposure in boars to di(2-ethylhexyl) phthalate (DEHP), a plasticizer reported to have toxic effects on rodent reproduction, would affect the sperm ability to undergo capacitation and acrosome reaction (AR) in vitro or give rise to a higher degree of chromatin instability associated with acid-induced denaturation. Spermatozoa were collected from 16 boars (n=8/group) 8-9 months of age, exposed to 300mg/kg body weight of DEHP or placebo per os three times a week, from 3 to 7 weeks of age. The spermatozoa were cryopreserved and examined post-thaw by flow cytometry for their ability to capacitate in vitro when exposed to the effector bicarbonate and to acrosome-react when exposed to calcium ionophores, using the lipid stain Merocyanine-540 (m-540), and peanut agglutinin-fluorescein isothiocyanate, respectively, as probes. The ability of the DNA to sustain denaturation in vitro was tested using a sperm chromatin structure assay (SCSA). No significant differences between the DEHP-exposed group and controls were found for any of the sperm attributes examined. Frozen-thawed spermatozoa showed similar rates of non-capacitated cells between groups, and were capacitated at similar rates. Rates of induced ARs were also similar. Values of DNA denaturation were low and showed no differences between groups. In conclusion, pre-pubertal exposure to DEHP does not seem, under the conditions of the present experiment, to affect the ability of frozen-thawed spermatozoa collected post-puberty to capacitate or acrosome-react (the main requisites for fertilization) or to present damage in their nuclear genome.