Sperm nuclei analysis and nuclear organization of a fertile boar-pig hybrid by 2d fish on both total and motile sperm fractions

A wide range of mammalian hybrids has recently been found by chance or through pop-ulation-screening programs, but studies about their fertilizing capacity remain scarce and incom-plete. Most of them are assumed to be sterile due to meiotic arrest caused by the failure of chromosome pairings. In this study, we evaluated both sperm meiotic segregation, by 2D fluorescent in situ hybridization (FISH) analysis, and sperm quality (Sperm Chromatin Structure Assay) by flow cy-tometer in a fertile boar-pig hybrid (2n = 37,XY) originating from a Nero Siciliano pig breed (Sus scrofa domesticus) and a wild boar (Sus scrofa ferus). Spermatozoa were also separated by a dual-layer (75-60%) discontinuous Percoll gradient, resulting in two fractions with a significantly better over-all quality in the motile sperm fraction. These data were confirmed by FISH analysis also, where the frequencies of spermatozoa with a regular chromosome composition were 27% in total sperm fraction and 64% in motile sperm fraction. We also evaluated the nuclear architecture in all counted spermatozoa, showing a chromatin distribution changing when chromosome abnormalities occur. Our results demonstrate that the chromosome pairing has a minimal effect on the sperm segregation and semen quality of a boar-pig hybrid, making it fertile and harmful for the conservation of au-tochthonous pig breeds.