title: Low amounts of mitochondrial reactive oxygen species define human sperm quality.
abstract: We have applied the mitochondria-specific superoxide fluorescent probe MitoSOX Red (MitoSOX) to detect mitochondria-specific reactive oxygen species (mROS) production in human sperm samples using flow cytometry. We show that human ejaculates are heterogeneous in terms of mROS production, with three subpopulations clearly detectable, comprising sperm that produce increasing amounts of mROS (MitoSOX-, MitoSOX+, and MitoSOX++). The sperm subpopulation producing the lowest amount of mROS represented the most functional subset of male gametes within the ejaculate, as it was correlated with the highest amount of live and non-apoptotic sperm and increased both in samples with better semen parameters and in samples processed by both density-gradient centrifugation and swim-up, both known to select for higher quality sperm. Importantly, the MitoSOX- subpopulation was clearly more prevalent in samples that gave rise to pregnancies following assisted reproduction. Our work, therefore, not only describe discreet human sperm heterogeneity at the mROS level but also suggests that mROS may represent a strategy to both evaluate sperm samples and isolate the most functional gametes for assisted reproduction.
authors.affiliation: Biology of Reproduction and Stem Cell GroupCNC-Center for Neuroscience and Cell Biology, University of Coimbra, 3004-517 Coimbra, PortugalHuman Reproduction ServiceUniversity Hospitals of Coimbra, Pr. Mota Pinto, Ed. São Jerónimo, 3000-075 Coimbra, PortugalPortuguese Institute for Blood and TransplantationCoimbra, PortugalFaculty of MedicineUniversity of Coimbra, Coimbra, PortugalDepartment of Life SciencesUniversity of Coimbra, PO Box 3046, 3001-401 Coimbra, PortugalBiology of Reproduction and Stem Cell GroupCNC-Center for Neuroscience and Cell Biology, University of Coimbra, 3004-517 Coimbra, PortugalHuman Reproduction ServiceUniversity Hospitals of Coimbra, Pr. Mota Pinto, Ed. São Jerónimo, 3000-075 Coimbra, PortugalPortuguese Institute for Blood and TransplantationCoimbra, PortugalFaculty of MedicineUniversity of Coimbra, Coimbra, PortugalDepartment of Life SciencesUniversity of Coimbra, PO Box 3046, 3001-401 Coimbra, Portugal.
authors.forename: Mónica
authors.initials: M
authors.lastname: Marques
authors.affiliation: Biology of Reproduction and Stem Cell GroupCNC-Center for Neuroscience and Cell Biology, University of Coimbra, 3004-517 Coimbra, PortugalHuman Reproduction ServiceUniversity Hospitals of Coimbra, Pr. Mota Pinto, Ed. São Jerónimo, 3000-075 Coimbra, PortugalPortuguese Institute for Blood and TransplantationCoimbra, PortugalFaculty of MedicineUniversity of Coimbra, Coimbra, PortugalDepartment of Life SciencesUniversity of Coimbra, PO Box 3046, 3001-401 Coimbra, PortugalBiology of Reproduction and Stem Cell GroupCNC-Center for Neuroscience and Cell Biology, University of Coimbra, 3004-517 Coimbra, PortugalHuman Reproduction ServiceUniversity Hospitals of Coimbra, Pr. Mota Pinto, Ed. São Jerónimo, 3000-075 Coimbra, PortugalPortuguese Institute for Blood and TransplantationCoimbra, PortugalFaculty of MedicineUniversity of Coimbra, Coimbra, PortugalDepartment of Life SciencesUniversity of Coimbra, PO Box 3046, 3001-401 Coimbra, Portugal.
authors.forename: Ana Paula
authors.initials: AP
authors.lastname: Sousa
authors.affiliation: Biology of Reproduction and Stem Cell GroupCNC-Center for Neuroscience and Cell Biology, University of Coimbra, 3004-517 Coimbra, PortugalHuman Reproduction ServiceUniversity Hospitals of Coimbra, Pr. Mota Pinto, Ed. São Jerónimo, 3000-075 Coimbra, PortugalPortuguese Institute for Blood and TransplantationCoimbra, PortugalFaculty of MedicineUniversity of Coimbra, Coimbra, PortugalDepartment of Life SciencesUniversity of Coimbra, PO Box 3046, 3001-401 Coimbra, Portugal.
authors.forename: Artur
authors.initials: A
authors.lastname: Paiva
authors.affiliation: Biology of Reproduction and Stem Cell GroupCNC-Center for Neuroscience and Cell Biology, University of Coimbra, 3004-517 Coimbra, PortugalHuman Reproduction ServiceUniversity Hospitals of Coimbra, Pr. Mota Pinto, Ed. São Jerónimo, 3000-075 Coimbra, PortugalPortuguese Institute for Blood and TransplantationCoimbra, PortugalFaculty of MedicineUniversity of Coimbra, Coimbra, PortugalDepartment of Life SciencesUniversity of Coimbra, PO Box 3046, 3001-401 Coimbra, PortugalBiology of Reproduction and Stem Cell GroupCNC-Center for Neuroscience and Cell Biology, University of Coimbra, 3004-517 Coimbra, PortugalHuman Reproduction ServiceUniversity Hospitals of Coimbra, Pr. Mota Pinto, Ed. São Jerónimo, 3000-075 Coimbra, PortugalPortuguese Institute for Blood and TransplantationCoimbra, PortugalFaculty of MedicineUniversity of Coimbra, Coimbra, PortugalDepartment of Life SciencesUniversity of Coimbra, PO Box 3046, 3001-401 Coimbra, Portugal.
authors.forename: Teresa
authors.initials: T
authors.lastname: Almeida-Santos
authors.affiliation: Biology of Reproduction and Stem Cell GroupCNC-Center for Neuroscience and Cell Biology, University of Coimbra, 3004-517 Coimbra, PortugalHuman Reproduction ServiceUniversity Hospitals of Coimbra, Pr. Mota Pinto, Ed. São Jerónimo, 3000-075 Coimbra, PortugalPortuguese Institute for Blood and TransplantationCoimbra, PortugalFaculty of MedicineUniversity of Coimbra, Coimbra, PortugalDepartment of Life SciencesUniversity of Coimbra, PO Box 3046, 3001-401 Coimbra, PortugalBiology of Reproduction and Stem Cell GroupCNC-Center for Neuroscience and Cell Biology, University of Coimbra, 3004-517 Coimbra, PortugalHuman Reproduction ServiceUniversity Hospitals of Coimbra, Pr. Mota Pinto, Ed. São Jerónimo, 3000-075 Coimbra, PortugalPortuguese Institute for Blood and TransplantationCoimbra, PortugalFaculty of MedicineUniversity of Coimbra, Coimbra, PortugalDepartment of Life SciencesUniversity of Coimbra, PO Box 3046, 3001-401 Coimbra, Portugal jramalho@ci.uc.pt.
authors.forename: João
authors.initials: J
authors.lastname: Ramalho-Santos
mesh_terms: D000328:Adult; D017209:Apoptosis; D015415:Biomarkers / Q000378:metabolism; D002470:Cell Survival; D005260:Female; D005434:Flow Cytometry; D006801:Humans; D008297:Male; D008928:Mitochondria / Q000378:metabolism*; D010617:Phenanthridines; D011247:Pregnancy; D018873:Pregnancy Rate; D017382:Reactive Oxygen Species / Q000378:metabolism*; D027724:Reproductive Techniques, Assisted; D055101:Semen Analysis / Q000379:methods; D013094:Spermatozoa / Q000378:metabolism* / Q000473:pathology
chemical_list: C521281:5-(6'-triphenylphosphoniumhexyl)-5,6-dihydro-6-phenyl-3,8-phenanthridinediammine; D015415:Biomarkers; D010617:Phenanthridines; D017382:Reactive Oxygen Species