Abstract

The aim of this study was to evaluate the effect of an acute exposure to a sinusoidal MF-ELF (50 Hz, 1mT) on the ability of boar mature spermatozoa to acquire the fertilizing competence in vitro. The spermatozoa exposed during the 4h of incubation to the MF-ELF were evaluated for morphological (surface morphology and acrosome integrity) and functional parameters (cell viability, motility, induction of acrosomal reaction, AR, and the ability to in vitro fertilize oocytes). In parallel, the intracellular Ca(2+) levels as well as the major mechanisms of Ca(2+) clearance were assessed: (45)Ca intakes and intracellular Ca(2+) sequestration by analyzing intracellular Ca(2+) elevation induced by thapsigargin or studying mitochondrial function with Mito-Tracker. The MF-ELF exposure did not affect sperm viability and morphology during the first h of incubation when sperm Ca(2+) homeostasis were already compromised. First of all, MF-ELF treated spermatozoa showed resting intracellular Ca(2+) levels significantly lower than those recorded in controls. This result was dependent on a lower extracellular Ca(2+) intake and from the inhibitory role exerted on both intracellular Ca(2+) storages. As a consequence, after 1h of incubation MF-ELF exposed cells displayed a reduced motility, a modest reactivity when coincubated with solubilized zonae pellucidae and a reduction in oocyte penetrating ability. After 2 or 4h of incubation, in addition, signs of morphological damage appeared on plasma membrane and at acrosomal level. In conclusion, MF-ELF influence negatively spermatozoa first by impairing cell Ca(2+) homeostasis then by dramatically affecting sperm morphology and function.