title: Intracytoplasmic sperm injection using DNA-fragmented sperm in mice negatively affects embryo-derived embryonic stem cells, reduces the fertility of male offspring and induces heritable changes in epialleles.
abstract: Intracytoplasmic sperm injection (ICSI) in mice using DNA-fragmented sperm (DFS) has been linked to an increased risk of genetic and epigenetic abnormalities both in embryos and offspring. This study examines: whether embryonic stem cells (ESCs) derived from DFS-ICSI embryos reflect the abnormalities observed in the DFS-ICSI progeny; the effect of DFS-ICSI on male fertility; and whether DFS-ICSI induces epigenetic changes that lead to a modified heritable phenotype. DFS-ICSI-produced embryos showed a low potential to generate ESC lines. However, these lines had normal karyotype accompanied by early gene expression alterations, though a normal expression pattern was observed after several passages. The fertility of males in the DFS-ICSI and control groups was compared by mating test. Sperm quantity, vaginal plug and pregnancy rates were significantly lower for the DFS-ICSI-produced males compared to in vivo-produced mice, while the number of females showing resorptions was higher. The epigenetic effects of DFS-ICSI were assessed by analyzing the phenotype rendered by the Axin1Fu allele, a locus that is highly sensitive to epigenetic perturbations. Oocytes were injected with spermatozoa from Axin1Fu/+ mice and the DFS-ICSI-generated embryos were transferred to females. A significantly higher proportion of pups expressed the active kinky-tail epiallele in the DFS-ICSI group than the controls.
  
  
  CONCLUSIONS
  1) ESCs cannot be used as a model of DFS-ICSI; 2) DFS-ICSI reduces sperm production and fertility in the male progeny; and 3) DFS-ICSI affects the postnatal expression of a defined epigenetically sensitive allele and this modification may be inherited across generations.
authors.affiliation: Dpto. de Reproducción Animal y Conservación de Recursos Zoogenéticos, Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria, Madrid, Spain.
authors.forename: Priscila
authors.initials: P
authors.lastname: Ramos-Ibeas
authors.affiliation: Dpto. de Reproducción Animal y Conservación de Recursos Zoogenéticos, Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria, Madrid, Spain.
authors.forename: Alexandra
authors.initials: A
authors.lastname: Calle
authors.affiliation: Dpto. de Reproducción Animal y Conservación de Recursos Zoogenéticos, Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria, Madrid, Spain.
authors.forename: Raúl
authors.initials: R
authors.lastname: Fernández-González
authors.affiliation: Dpto. de Reproducción Animal y Conservación de Recursos Zoogenéticos, Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria, Madrid, Spain.
authors.forename: Ricardo
authors.initials: R
authors.lastname: Laguna-Barraza
authors.affiliation: Dpto. de Reproducción Animal y Conservación de Recursos Zoogenéticos, Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria, Madrid, Spain.
authors.forename: Eva
authors.initials: E
authors.lastname: Pericuesta
authors.affiliation: Dpto. de Reproducción Animal y Conservación de Recursos Zoogenéticos, Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria, Madrid, Spain.
authors.forename: Antonia
authors.initials: A
authors.lastname: Calero
authors.affiliation: Dpto. de Reproducción Animal y Conservación de Recursos Zoogenéticos, Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria, Madrid, Spain.
authors.forename: Miguel Ángel
authors.initials: MÁ
authors.lastname: Ramírez
authors.affiliation: Dpto. de Reproducción Animal y Conservación de Recursos Zoogenéticos, Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria, Madrid, Spain.
authors.forename: Alfonso
authors.initials: A
authors.lastname: Gutiérrez-Adán
mesh_terms: D000483:Alleles; D000818:Animals; D053938:DNA Fragmentation; D005260:Female; D008297:Male; D051379:Mice; D011247:Pregnancy; D020554:Sperm Injections, Intracytoplasmic / Q000009:adverse effects*; D013094:Spermatozoa / Q000378:metabolism / Q000502:physiology*
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